The development and implementation of pathological parameters and molecular testing impact prognosis of colorectal adenocarcinoma

Objective:This study aims to analyze how changes in pathological diagnosis practice and molecular detection technology have affected clinical outcomes for colorectal cancer(CRC)patients in Fudan University Shanghai Cancer Center(FUSCC).Methods:This retrospective cohort study analyzed 21,141 pathologically confirmed CRC cases diagnosed at FUSCC from 2008 to 2020.Patients were divided into five groups for different analytical purposes:(1)the before vs.since 2014 groups to analyze the influence of the changes in the classification criteria of pT3 and pT4 staging on the survival of patients;(2)the partial vs.total mesorectal excision(TME)groups to analyze whether evaluation of completeness of the mesorectum have impact on the survival of patients;(3)the tumor deposit(TD)(+)N0 vs.TD(+)N1c groups to analyze the influence of the changes in the pN staging on the survival of patients with positive TD and negative regional lymph node metastasis(LNM);(4)the before vs.since 2013 groups to analyze the influence of the changes in the testing process of deficient mismatch repair on the survival of patients;and(5)the groups with vs.without RAS/BRAF gene mutation testing to analyze the influence of these testing on the survival of patients.Patients’clinicopathological parameters,including age at diagnosis,sex,tumor size,location,differentiation,mucinous subtype,TD,lymphovascular invasion,perineural invasion,tumor depth,LNM and distant metastasis,and tumor-node-metastasis(TNM)stage,were compared between groups.Kaplan-Meier analysis with log rank method was performed for patients’overall survival(OS)and disease-free survival(DFS)analyses.Results:In pathological reports,there were three parameter changes that impacted patient outcomes.Firstly,changes in the pT staging criteria led to a shift of the ratio of patients with stage pT3 to stage pT4 from 1:110.9 to 1:0.26.In comparison to patients admitted before 2014(n=4,754),a significant difference in prognosis between pT3 and pT4 stages was observed since 2014(n=9,965).Secondly,we began to evaluate the completeness of the mesorectum since 2016.As a result,91.0%of patients with low rectal cancer underwent TME(n=4,111)surgery,and patients with TME had significantly better OS compared with partial mesorectal excision(PME,n=409).Thirdly,we began to stage TD(+)LNM(-)as N1c since 2017.The results showed that N1c(n=127)but not N0(n=39)can improve the prognosis of patients without LNM and distal metastasis.In molecular testing,there have been three and five iterations of updates regarding mismatch repair(MMR)/microsatellite instability(MSI)status and RAS/BRAF gene mutation detection,respectively.The standardization of MMR status testing has sharply decreased the proportion of deficient MMR(dMMR)patients(from 32.5%to 7.4%)since 2013.The prognosis of patients underwent MMR status testing since 2013(n=867)were significantly better than patients before 2013(n=1,313).In addition,detection of RAS/BRAF gene mutation status(n=5,041)resulted in better DFS but not OS,for patients with stage I-III disease(n=16,557).Conclusion:Over the past few decades,updates in elements in pathological reports,as well as the development of standardized tests for MMR/MSI status and RAS/BRAF gene mutations have significantly improved patient outcomes.

Cytokeratms and carcinoembryonic antigen in diagnosis,staging and prognosis of colorectal adenocarcinoma

AIM: To evaluate the serum levels of cytokeratins and carcinoembryonic antigen (CEA) in diagnosis, staging and prognosis of patients with colorectal adenocarcinoma.METHODS: The sample consisted of 169 patients. One hundred blood donors formed the control group. Radical surgery was performed on 120 patients, with an average follow-up duration of 22.3 mo. Relapses occurred in 23individuals after an average of 18.09 mo. CEA was assayed via the Delfia(R) method with a limit of 5 ng/mL. Cytokeratins were assayed via the LIA-mat(R) TPA-M Prolifigen(R) method with a limit of 72 U/L.RESULTS: In the diagnosis of patients with colorectal adenocarcinoma, CEA showed a sensitivity of 56%, a specificity of 95%, a positive predictive value of 94%, a negative predictive value of 50% and an accuracy of 76.8%.TPA-M had a sensitivity of 70%, a specificity of 96%, a positive predictive value of 97%, a negative predictive value of 66% and an accuracy of 93.6%. The elevation of one of the markers was shown to have a sensitivity of 76.9%, a specificity of 91%, a positive predictive value of 93.5%, a negative predictive value of 70% and an accuracy of 83.6%.There was no variation in the levels of the markers according to the degree of cell differentiation while there was an elevation in their concentrations in accordance with the increase in neoplastic dissemination. There was a statistically significant difference between the patients with stage Ⅳ lesions and those with stages Ⅰ, Ⅱ and Ⅲ tumors.With regard to CEA, the averagelevel was 14.2 ng/mL in patients with stage Ⅰ lesions, 8.5 ng/mL in patients with stage Ⅱ lesions, 8.0 ng/mL in patients with stage Ⅲ lesions and 87.7 ng/mL in patients with stage Ⅳ lesions. In relation to TPA-M, the levels were 153.1 U/L in patients with stage Ⅰtumors, 106.5 U/L in patients with stage Ⅱ tumors, 136.3 U/L in patients with stage Ⅲ tumors and 464.3 U/L in patients with stage Ⅳ tumors. There was a statistical difference in patients with a high CEA level in relation to a shorter survival(P＜0.05). However, there was no correlation between patients with high TPA-M levels and prognostic indices of patients undergoing radical surgery.CONCLUSION: Cytokeratins demonstrate a greater sensitivity than CEA in the diagnosis of colorectal adenocarcinoma.There is an increase in the sensitivity of the markers with tumor dissemination. Cytokeratins cannot identify the worse prognosis in patients undergoing radical surgery.Cytokeratins constitute an advance in the direction of a perfect tumor marker in the treatment of patients with colorectal cancer.

Cancer stem cell markers CD133 and CD24 correlate with invasiveness and differentiation in colorectal adenocarcinoma

AIM:To verify that CD markers are available for detecting cancer stem cell populations and to evaluate their clinical significance in colon cancer.METHODS:Immunohistochemistry for CD133,CD24 and CD44 was performed on the tissue microarray of 523 colorectal adenocarcinomas.Medical records were reviewed and clinicopathological analysis was performed.RESULTS:In colorectal adenocarcinoma,128 of 523 cases(24.5%) were positive and 395 cases(75.5%) were negative for CD133 expression.Two hundred and sixty-four of 523 cases(50.5%) were positive and 259 cases(49.5%) were negative for CD24 expression.Five hundred and two of 523 cases(96%) were negative and 21 cases(4%) were positive for CD44 expression.Upon clinicopathological analysis,CD133 expression was present more in male patients(P = 0.002) and in advanced T stage cancer(P = 0.024).Correlation between CD24 expression and clinicopathological factors was seen in the degree of differentiation(P = 0.006).Correlation between CD44 expression and clinicopathological factors was seen in the tumor size(P = 0.001).Survival was not significantly related to CD133,CD24 and CD44 expression.CONCLUSION:CD markers were related to invasiveness and differentiation of colorectal adenocarcinoma.However,CD expression was not closely related to survival.

Sonic hedgehog-Gli1 pathway in colorectal adenocarcinomas

AIM： To determine the role of Sonic hedgehog （Shh） pathway in colorectal adenocarcinomas through analysis of the expression of Shh pathway-related molecules, Shh, Ptchl, hedgehog-interacting protein （Hip）, Gil1, Gli3 and PDGFRα. METHODS： Expression of Shh in 25 colorectal adenocarcinomas was detected by RT-PCR,in situ hybridization and immunohistochemistry. Expression of Ptchl was observed by in situ hybridization and immunohistochemistry. Expression of Hip, Gil1, Gli3 and PDGFRα was analyzed by in situ hybridization. RESULTS： Expression of cytokeratin AE1/AE3 was observed in the cytoplasm of colorectal crypts. Members of the Hh signaling pathway were expressed in colorectal epithelium. Shh was expressed in cytoplasm of dysplastic epithelial cells, while expression of Ptchl, Hip and Gill were mainly detected in the malignant crypts of adenocarcinomas. In contrast, PDGFRα was expressed highly in aberrant crypts and moderately in the stroma. Expression of Gli3 could not be detected in colorectal adenocarcinomas. CONCLUSION： These data suggest that Shh-Ptchl-Gli1 signaling pathway may play a role in the progression of colorectal tumor.

Relationship between loss of heterozygosity of microsatellite on DCC gene and prognosis of colorectal adenocarcinoma

AIM To investigate the relationship between the loss of heterozygosity (LOH) of microsatellite on the Deleted in colorectal carcinoma (DCC) gene and the prognosis of colorectal adenocarcinoma. METHODS A retrospective study of 58 cases of colorectal adenocarcinoma with the follow up data and paired control normal mucosal tissues from 1983 to 1985 from files of the Department of Pathology, West China University of Medical Sciences was carried out by PCR microsatellite method. Tumors were graded as well, moderately and poorly differentiated in 16, 35 and 7 cases, and staged as Dukes′ A, B, C in 11, 30 and 17 cases, respectively. RESULTS Heterozygosity of microsatellite was lost on DCC gene in 18 of 58 (31 0%) cases of colorectal adenocarcinoma. The 5 year survival rate between the positive and negative for LOH was 44 4% and 77 5%, respectively, being significantly different ( P <0 05). It was suggested that the LOH on microsatellite of DCC gene was correlated to the prognosis, but not to the differentiation ( P >0 05) and Dukes′ stage ( P >0 05) in colorectal adenocarcinoma. CONCLUSION LOH of microsatellite on DCC gene may be one of malignant markers. Combined with the traditional prognostic indices, LOH can predict the prognosis of colorectal adenocarcinoma.

Survivin isoforms and clinicopathological characteristics in colorectal adenocarcinomas using real-time qPCR

AIM:To investigate three isoforms of survivin in colorectal adenocarcinomas.METHODS:We used the LightCycler Technology(Roche),along with a common forward primer and reverse primers specific for the splice variants and two common hybridization probes labeled with fluorescein and LightCycler-Red fluorophore(LC-Red 640).Real time quantitative polymerase chain reaction(PCR) was performed on cDNAs from 52 tumor specimens from colorectal cancer patients and 10 unrelated normal colorectal tissues.In the patients group,carcinoembryonic antigen(CEA) and CA19-9 tumor markers were also measured immunochemically.RESULTS:Wild type survivin mRNA isoform was expressed in 48%of the 52 tumor samples,survivin-2b in 38%and survivin-ΔΕx3 in 29%,while no expression was found in normal tissues.The mRNA expression of wild type survivin presented a significant correlation with the expression of the ratio of survivin-2b,survivin-ΔΕx3,survivin-2b/wild type survivin and survivin-ΔΕx3/wild type survivin(P<0.001).The mRNA expression of wildsurvivin and survivin-ΔΕx3 was related with tumor size and invasion(P=0.006 and P<0.005,respectively).A significant difference was found between survivin-2b and morphologic cancer type.Also,the ratio of survivin-ΔEx3/ wild-survivin was significantly associated with prognosis.No association was observed between the three isoforms and grade,metastasis,Dukes stage and gender.The three isoforms were not correlated with CEA and CA19-9.CONCLUSION:Survivin isoforms may play a role in cell apoptosis and their quantification could provide information about clinical management of patients suffering from colorectal cancer.

TSPAN1 protein expression:A significant prognostic indicator for patients with colorectal adenocarcinoma

AIM:To determine if TSPAN1 overexpression is associated with clinicopathological and prognostic factors in human colorectal adenocarcinoma.METHODS:Total RNA was extracted in 20 human adenocarcinoma tissues for TSPAN1 mRNA assay by RT-PCR.Eighty-eight specimens of human colorectal adenocarcinoma were surgically removed.TSPAN1 protein levels in cancer tissues were determined by immunohistochemistry using a polyclonal antibody against self-prepared TSPAN1.The correlation between TSPAN1 expression and the clinicopathological factors and the overall survival rate was analyzed by univariate and multivariate assay.RESULTS:TSPAN1 mRNA was detected in 90.0%(18/20) of cancerous tissues.The light density of TSPAN1 mRNA expression levels was 0.89 ± 0.30 in adenocarcinoma by gel-image system.TSPAN1 protein expression was detected in 78.41%(69/88) and weakly expressed in 40% normal colorectal tissues.There were significant differences between colorectal adenocarcinoma and normal control epithelium(P < 0.05).TSPAN1 protein expression in colorectal cancerous tissue was significantly correlated with the histological grade,cell expression PCNA,lymph nodal metastasis and TNM staging of the disease.Patients with TSPAN1 protein overexpression had a significantly shorter survival period than that in patients with TSPAN1 protein negative or weak expression,respectively(P < 0.05).Furthermore,by multivariate analysis,TSPAN1 protein expression demonstrated an independent prognostic factor for human colorectal cancers(P < 0.05,relative risk 0.755;95% confidence interval 0.302-1.208).CONCLUSION:The expression of TSPAN1 gene is increased in colorectal carcinoma,suggesting that TSPAN1 might serve as an independent prognostic factor for the colorectal adenocarcinoma patients.

Identification of differently expressed genes in human colorectal adenocarcinoma

AIM： To investigate the differently expressed genes in human colorectal adenocarcinorna.METHODS： The integrated approach for gene expression profiling that couples suppression subtractive hybridization, high-throughput cDNA array, sequencing, bioinformatics analysis, and reverse transcriptase real- time quantitative polymerase chain reaction （PCR） was carried out. A set of cDNA clones including 1260 SSH inserts amplified by PCR was arrayed using robotic printing. The cDNA arrays were hybridized with florescent-labeled probes prepared from RNA of human colorectal adenocarcinoma （HCRAC） and normal colorectal tissues.RESULTS： A total of 86 genes were identified, 16 unknown genes and 70 known genes. The transcription factor Sox9 influencing cell differentiation was downregulated. At the same time, Heat shock protein 10 KDis downregulated and Calmoulin is up-regulated.CONCLUSION： Downregulation of heat shock protein 10 KD lost its inhibition of Ras, and men attenuated the Ras GTPase signaling pathway, increased cell proliferation and inhibited cell apoptosis. Down-regulated transcription factor Sox9 influences cell differentiation and cell-specific gene expression. Down-regulated Sox9 also decreases its binding to calmodulin, accumulates calmodulin as receptor-activated kinase and phosphorylase kinase due to the activation of PhK.

Mucinous adenocarcinoma:A unique clinicopathological subtype in colorectal cancer

Mucinous adenocarcinoma(MAC)is a unique clinicopathological subtype of colorectal cancer,which is characterized by extracellular mucinous components that comprise at least 50%of the tumor tissue.The clinical characteristics,molecular features,response to chemo-/radiotherapy,and prognosis of MAC are different from that of non-MAC(NMAC).MAC is more common in the proximal colon,with larger volume,higher T-stage,a higher proportion of positive lymph nodes,poorer tumor differentiation,and a higher proportion of peritoneal implants compared to NMAC.Although biopsy is the main diagnostic method for MAC,magnetic resonance imaging is superior in accuracy,especially for rectal carcinoma.The aberrant expression of mucins,including MUC1,MUC2 and MUC5AC,is a notable feature of MAC,which may be related to tumor invasion,metastasis,inhibition of apoptosis,and chemo-/radiotherapy resistance.The genetic origin of MAC is mainly related to BRAF mutation,microsatellite instability,and the CpG island methylator phenotype pathway.In addition,the poor prognosis of rectal MAC has been confirmed by various studies,and that of colonic MAC is still controversial.In this review,we summarize the epidemiology,clinicopathological characteristics,molecular features,methods of diagnosis,and treatments of MAC in order to provide references for further fundamental and clinical research.

Micronucleus analysis in patients with colorectal adenocarcinoma and colorectal polyps

AIM:To determine,by counting micronucleus (MN) frequencies,whether chromosomal or DNA damage have an effect on the pathogenesis of early colorectal adenocarcinoma (CRC). METHODS:We analyzed MN frequencies in 21 patients with CRC,24 patients with colon polyps [10 neoplastic polyps (NP) and 14 non-neoplastic polyps (NNP)] and 20 normal controls. RESULTS:MN frequency was significantly increased in CRC patients and in NP patients compared with controls (3.72 ± 1.34,3.58 ± 1.21 vs 1.97 ± 0.81,P < 0.001). However,there was no difference in the MN frequency between CRC patients and NP patients (P > 0.05). Similarly,there was no difference in the MN frequency between NNP patients (2.06 ± 0.85) and controls (P > 0.05). CONCLUSION:Our results suggest increased chromosome/DNA instabilities may be associated with the pathogenesis of early CRC.

B2 adrenergic receptors and morphological changes of the enteric nervous system in colorectal adenocarcinoma

To study the morphology of the enteric nervous system and the expression of beta-2 adrenergic (B2A) receptors in primary colorectal cancer.METHODSIn this study, we included forty-eight patients with primary colorectal cancer and nine patients for control tissue from the excision of a colonic segment for benign conditions. We determined the clinicopathological features and evaluated the immunohistochemical expression pattern of B2A receptors as well as the morphological changes of the enteric nervous system (ENS). In order to assess statistical differences, we used the student t-test for comparing the means of two groups and one-way analysis of variance with Bonferroni’s post hoc analysis for comparing the means of more than two groups. Correlations were assessed using the Pearson’s correlation coefficient.RESULTSB2A receptors were significantly associated with tumor grading, tumor size, tumor invasion, lymph node metastasis (P < 0.05), while there were no statistically significant associations with gender, CRC location and gross appearance (P > 0.05). We observed, on one hand, a decrease of the relative area for both Auerbach and Meissner plexuses with the increase of the tumor grading, and on the other hand, an increase of the relative area of other nervous elements not in the Meissner plexus or in the Auerbach plexus with the tumor grading. For G1 tumors we found that epithelial B2A area showed an inverse correlation with the Auerbach plexus areas [r(14) = -0.531, P < 0.05], while for G2 tumors, epithelial B2A areas showed an indirect variation with both the Auerbach plexus areas [r(14) = -0.453, P < 0.05] and the Meissner areas [r(14) = -0.825, P < 0.01]. For G3 tumors, the inverse dependence increased for both Auerbach [r(14) = -0.587, P < 0.05] and Meissner [r(14) = -0.934, P < 0.05] plexuses.CONCLUSIONB2A receptors play an important role in colorectal carcinogenesis and can be utilized as prognostic factors. Furthermore, study of the ENS in colorectal cancer may lead to targeted molecular therapies.

Characteristics and risk factor analyses of high-grade intraepithelial neoplasia in older patients with colorectal polyps

BACKGROUND According to the degree of intradermal neoplasia in the colorectal exhalation,it can be divided into two grades:Low-grade intraepithelial neoplasia(LGIN)and high-grade intraepithelial neoplasia(HGIN).Currently,it is difficult to accurately diagnose LGIN and HGIN through imaging,and clinical diagnosis depends on postoperative histopathological diagnosis.A more accurate method for evaluating HGIN preoperatively is urgently needed in the surgical treatment and nursing intervention of colorectal polyps.AIM To explore the characteristics and risk factors of HGIN in older patients with colorectal polyps.METHODS We selected 84 older patients diagnosed with HGIN as the HGIN group(n=95 colonic polyps)and 112 older patients diagnosed with LGIN as the LGIN group(n=132 colonic polyps)from Shandong Provincial Hospital Affiliated to Shandong First Medical University.The endoscopic features,demographic characteristics,and clinical manifestations of the two patient groups were compared,and a logistic regression model was used to analyze the risk factors for HGIN in these patients.RESULTS The HGIN group was older and had a higher number of sigmoid colon polyps,rectal polyps,pedunculated polyps,polyps≥1.0 cm in size,polyps with surface congestion,polyps with surface depression,and polyps with villous/tubular adenomas,a higher proportion of patients with diabetes and a family history of colorectal cancer,patients who experienced rectal bleeding or occult blood,patients with elevated carcinoembryonic antigen(CEA)and cancer antigen 199(CA199),and lower nutritional levels and higher frailty levels.The polyp location(in the sigmoid colon or rectum),polyp diameter(≥1.0 cm),pathological diagnosis of(villous/tubular adenoma),family history of colorectal cancer,rectal bleeding or occult blood,elevated serum CEA and CA199 levels,lower nutritional levels and higher frailty levels also are independent risk factors for HGIN.CONCLUSION The occurrence of high-grade neoplastic transformation in colorectal polyps is closely associated with their location,size,villous/tubular characteristics,family history,elevated levels of tumor markers,and lower nutritional levels and higher frailty levels.

Protein Expression of STAT3, pSTAT3, MMP-7 and VEGF in Colorectal Adenocarcinoma: An Immunohistochemical Study

Background: The purpose of the present study is to investigate the expression levels of STAT3, pSTAT3, MMP-7 and VEGF in colorectal adenocarcinoma, and also to determine association with the clinico-pathological parameters and?co-expression of these genes. Methods: An immunohistochemical method was used to evaluate the expression of MMP-7 and VEGF genes in 93 archival tissues whereas STAT3 and pSTAT3 expression was determined in 75 cases. Results: Overexpression of STAT3 was detected in 26.7% (20/75), pSTAT3 in 13.4% (10/75), MMP-7 in 38.8% (36/93) and VEGF in 59.2% (55/93) of the colorectal carcinomas. STAT3, MMP-7 and VEGF immunopositivity were significantly correlated with poorly-differentiated tumors (P = 0.004;P = 0.03;P = 0.002, respectively) but not with other parameters. However, pSTAT3 immunostaining was not significantly associated with the clinico-pathological characteristics. Significant relationship was noted between overexpression of pSTAT3 and STAT3 (P < 0.001), pSTAT3 and VEGF (P = 0.044), pSTAT3 and MMP-7 (P = 0.003), and STAT3 and VEGF (P = 0.037) but marginal association was detected between STAT3 and MMP-7 (P = 0.057), and MMP-7 and VEGF (P = 0.052). Conclusion:?Our data suggest that expression of these genes?may have an important role in tumor dedifferentiation and?may be useful as indicators of biologic aggressiveness. Co-expression of the biomarkers by cancer cells?may have important implications in colorectal cancer biology and?could be useful biological markers of the malignant phenotype.

Skeletal muscle metastasis from colorectal adenocarcinoma:A literature review

BACKGROUND Colorectal adenocarcinoma is the third most common cancer worldwide.It accounts for almost 10%of all cancer-related deaths.Skeletal muscle is a very unusual site for metastasis from colorectal cancers and is associated with a poor prognosis and high mortality.AIM To review the literature for cases of skeletal muscle metastasis(SMM)from colorectal adenocarcinoma.METHODS A systematic literature search using a validated search strategy was carried out to identify the incidence of SMM associated with colorectal adenocarcinoma.The studies identified were tabulated in a PRISMA,and data was extracted in a tabulated form.RESULTS Twenty-nine studies were included in this literature review.SMM was most commonly detected in the thigh muscles.Most of the tumours had originated from the rectum or the right colon.The histopathology of the primary tumour was generally advanced.The mean time interval between the primary tumour and onset of SMM was 22 mo.In 3 cases,asymptomatic SMM had been picked up by advanced imaging systems,like fluorodeoxyglucose-positron emission tomography scan.CONCLUSION SMM from colorectal adenocarcinomas is a rare complication.However,it is possible that the low incidence could be due to under-reporting.Early use of advanced imaging techniques and a high index of clinical suspicion might increase the reporting of SMM from colorectal adenocarcinoma.

THE FURTHER STUDY ON BIOLOGICAL BEHAVIOR OF COLORECTAL ADENOCARCINOMA:THE EXTRACELLULAR PROTEOGLYCAN DEGRADED BY ARYLSULFATASE B

The fresh tissues were obtained from 64 colorectal adenocarcinoma (43 well-differentiated and moderately-differentiated adenocarcinoma, 12 poorly- differentiated adenocarcinoma and 9 mutinous cell carcinoma including signet ring cell carcinoma) during surgical operation. The resected edge of each specimen was used for control group. The arylsulfatase B was studied by histochentical staining in different types of colorectal adenocarcinoma, among which 19 cases were investigated by electronhistochemical staining so as to observe the Ruthenium Red granules alteration which represented the extracellular proteoglycan changes and ultrastructure of cancer cells.The results showed that the mutinous cell carcinoma was of the most Intensive arylsulfatase B activity and has a lot of secretory granules with various electron densities in the cytoplasm. The Ruthenium Red granules close to the cancer cell disappeared, a part of remainders changed into the lowered electron density and indistinct shape. In contrast, the other types adenocarcinoma revealed less enzyme activity and a fewer secretory granules. The Ruthenium Red granules near the cancer nest showed that their electron density and size were identical with those of the control group. All of these mentioned above indicate that mucinouscell carcinoma may release hydrolase into pericancerous matrix to degrade the proteoglycans. In view of the network structure formed by proteoglycan in the connective tissue, network has ability to hinder the cancer cell spreading. Because the arylsulfatase B is able to degrade the dermatan sulfate proteoglycan which is component of proteoglycans in the extracellular matrix of human colon. We consider that the arylsulfatase B may lead to destruction of the network barriers in the connective tissue in favour of cancer cell invasion. So the mucinous cell carcinoma is more malignant than those of other colorectal adenocarcinoma.

CORRELATIONSHIP BETWEEN CELLULAR DNA AND AgNOR PROTEIN CONTENT IN THE DEVELOPING COURSE OF COLORECTAL ADENOCARCINOMA

Cellular NDA and AgNOR Protein contents were evaluated byautomatic image analysis in tissue sections stained hy combined Feulgen-AgNOR staining method in 9 normal colonic mucosae, 45 colorectal adenomas and 27 adenocarcinomas. The results indicated that during the course that the normal colonical mucosa developed to colorectal adenocarcinoma via adenoma the DNA and AgNOR protein contents increased gradually and there were very significant correlationships between the DNA and the AgNOR protein contents of adenoma group and adenocarcinoma group. However, there were considerahle overlaping in the DNA or AgNOR Protein content and considerahle overlaping cases between adenoma and normal colonic mucosa groups and between adenoma and adenocarcinoma groups. But the overlaping scope in NDA and AgNOR Protein content and the number of overlaping cases were reduced significantly by assessing the correlationship between the DNA and AgNOR protein content. Therefore, it is much more reliable to distinguish colorectal adenomas from adnocarcinomas by using the correlationship between the cellular DNA and the AgNOR Protein contents in the same specimens.

Diagnostic usefulness of selected proteases and acute phase factors in patients with colorectal adenocarcinoma

BACKGROUND Uncontrolled growth and loss of control over basic metabolic functions,leading to invasive proliferation and metastases,are the salient traits of malignant tumors in general and colorectal cancer in particular.Invasion and metastases hinder effective tumor treatment.While surgical techniques and radiotherapy can be used to remove tumor focus,only chemotherapy can eliminate dispersed neoplastic cells.However,the efficacy of the latter method is limited in the advanced stages of the disease.Therefore,recognition of the mechanisms involved in neoplastic cell spreading is indispensable for developing effective therapies.AIM To use a number of biomarkers involved in cancer progression and identify a panel that could be used for effective early diagnosis.METHODS We recruited 185 patients with colorectal adenocarcinoma(98 men,87 women with median age 63).Thirty-five healthy controls were sex and age-matched.Dukes’staging was as follows:A=22,B=52,C=72,D=39.We analyzed patients'blood serum before surgery.We determined:(1)Cathepsin B(CB)with Barrett's method(fluorogenic substrate);(2)Leukocytic elastase(LE)in a complex with alpha 1 trypsin inhibitor(AAT)using the immunoenzymatic MERCK test;(3)Total sialic acid(TSA)with the colorimetric periodate-resorcinol method;(4)Lipid-bound sialic acid(LASA)with the colorimetric Taut's method;and(5)The antitrypsin activity(ATA)employing the colorimetric test.RESULTS In patients,the values of the five biochemical parameters were as follows:CB=16.1±8.8 mU/L,LE=875±598μg/L,TSA=99±31 mg%,LASA=0.68±0.33 mg%,and ATA=3211±1504 U/mL.Except for LASA,they were significantly greater than those of controls:CB=11.4±6.5 mU/L,LE=379±187μg/L,TSA=71.4±15.1 mg%,LASA=0.69±0.28 mg%,and ATA=2016±690 U/mL.For CB and LASA,the differences between the four Dukes’stages and controls were not statistically significant.The inter-stage differences for CB and LASA were also absent.The receiver operating characteristic(ROC)analysis revealed the potential diagnostic value of CB,TSA,and ATA.The area under ROC,sensitivity,and specificity for these three parameters were:0.85,72%,90%;0.75,66%,77%;and 0.77,63%,84%,respectively.The sensitivity and specificity for the threeparameter panel CB-TSA-ATA were equal to 88.2%and 100%,respectively.CONCLUSION The increased value of CB,TSA,and ATA parameters are associated with tumor biology,invasion,and metastasis of colorectal cancer.The presented evidence suggests the potential value of the CB-TSA-ATA biochemical marker panel in early diagnostics.

Etoposide sensitizes CT26 colorectal adenocarcinoma to radiation therapy in BALB/c mice

AIM： To investigate the combined effect of etoposide and radiation on CT26 colorectal adenocarcinoma implanted into BALB/c mice. METHODS： We evaluated the radiosensitizing effect of etoposide on CT26 colorectal adenocarcinoma in a syngeneic animal model. BALB/c mice were subcutaneously implanted with CT26 cells and divided into four groups： Gonlyol （intra-peritoneal saline×2） group, etoposide （5 mg/kg intra-peritoneally×2） group, radiation therapy （RT 5 Gy×2 fractions） group, and combination therapy with etoposide （5 mg/kg intra-peritoneally 1 h before radiation） group. RESULTS： Tumor growth was significantly inhibited by RT and combination therapy. The effect of combination therapy was better than that of RT. No significant changes were noted in body weight, plasma alanine aminotransferase, or creatinine in any group. The leukocyte count significantly but transiently decreased in the RT and combination therapy groups, but not in the etoposide and control groups. There was no skin change or hair loss in the RT and combination therapy groups. CONCLUSION： Etoposide can sensitize CT26 colorectal adenocarcinoma in BALB/c mice to RT without significant toxicity.

Somatic mutations in FAT cadherin family members constitute an underrecognized subtype of colorectal adenocarcinoma with unique clinicopathologic features

BACKGROUND The FAT cadherin family members(FAT1,FAT2,FAT3 and FAT4)are conserved tumor suppressors that are recurrently mutated in several types of human cancers,including colorectal carcinoma(CRC).AIM To characterize the clinicopathologic features of CRC patients with somatic mutations in FAT cadherin family members.METHODS We analyzed 526 CRC cases from The Cancer Genome Atlas PanCancer Atlas dataset.CRC samples were subclassified into 2 groups based on the presence or absence of somatic mutations in FAT1,FAT2,FAT3 and FAT4.Individual clinicopathological data were collected after digital slide review.Statistical analysis was performed using t tests and chi-square tests.RESULTS This CRC study cohort had frequent mutations in the FAT1(10.5%),FAT2(11.2%),FAT3(15.4%)and FAT4(23.4%)genes.Two hundred CRC patients(38.0%)harbored somatic mutations in one or more of the FAT family genes and were grouped into the FAT mutated CRC subtype.The FAT-mutated CRC subtype was more commonly located on the right side of the colon(51.0%)than in the rest of the cohort(30.1%,P<0.001).It showed favorable clinicopathologic features,including a lower rate of positive lymph nodes(pN1-2:33.5%vs 46.4%,P=0.005),a lower rate of metastasis to another site or organ(pM1:7.5%vs 16.3%,P=0.006),and a trend toward an early tumor stage(pT1-2:25.0%vs 18.7%,P=0.093).FAT somatic mutations were significantly enriched in microsatellite instability CRC(28.0%vs 2.1%,P<0.001).However,FAT somatic mutations in microsatellite stable CRC demonstrated similar clinicopathologic behaviors,as well as a trend of a better diseasefree survival rate(hazard ratio=0.539;95%confidence interval:0.301-0.967;log-rank P=0.073).CONCLUSION FAT cadherin family genes are frequently mutated in CRC,and their mutation profile defines a subtype of CRC with favorable clinicopathologic characteristics.

Colorectal adenocarcinoma patients with M1a diseases gain more clinical benefits from palliative primary tumor resection than those with M1b diseases: A propensity score matching analysis

BACKGROUND Surgical resection is regarded as the only potentially curative treatment option for patients with metastatic colorectal cancer(CRC).The National Comprehensive Cancer Network clinical practice guidelines do not recommend palliative surgery unless there is a risk of severe symptoms.However,accumulating evidence has shown that palliative surgery is associated with more favorable outcomes for patients with metastatic CRC.AIM To investigate the separate role of palliative primary tumor resection for patients with stage IVA(M1a diseases)and stage IVB(M1b diseases)colorectal adenocarcinoma(CRA).METHODS CRA patients diagnosed from 2010 to 2015 with definite M1a and M1b categories according to the 8th edition of American Joint Committee on Cancer staging system were selected from the Surveillance Epidemiology and End Results(SEER)database.To minimize potential selection bias,the data were adjusted by propensity score matching(PSM).Baseline characteristics,including gender,year of diagnosis,age,marital status,primary site,surgical information,race,grade,chemotherapy,and radiotherapy,were recorded and analyzed.Univariate and multivariate analyses were performed to explore the separate role of palliative surgery for patients with M1a and M1b diseases.RESULTS A total of 19680 patients with metastatic CRA were collected from the SEER database,including 10399 cases of M1a diseases and 9281 cases of M1b diseases.Common independent prognostic factors for both M1a and M1b patients included year of diagnosis,age,race,marital status,primary site,grade,surgery,and chemotherapy.After PSM adjustment,3732 and 3568 matched patients in the M1a and M1b groups were included,respectively.Patients receiving palliative primary tumor resection had longer survival time than those without surgery(P<0.001).For patients with M1a diseases,palliative resection could increase the median survival time by 9 mo;for patients with M1b diseases,palliative resection could prolong the median survival time by 7 mo.For M1a diseases,patients with lung metastasis had more clinical benefit from palliative resection than those with liver metastasis(15 mo for lung metastasis vs 8 mo for liver metastasis,P<0.001).CONCLUSION CRA patients with M1a diseases gain more clinical benefits from palliative primary tumor resection than those with M1b diseases.Those patients with M1a(lung metastasis)have superior long-term outcomes after palliative primary tumor resection.