Effect of tumor suppressor in lung cancer-1 on growth inhibition of MG63 cell line

Objective： The aim of this study was to establish the osteosarcoma cell sublines which stably expressing tumor suppressor in lung cancer-1 （TSLC1） gene and evaluate its effect on growth inhibition of human osteosarcoma cell line MG63. Methods： The recombinant plasmid pCI-TSLC1 was stably transfected into MG63 cells with Lipofectamine 2000. The posi- tive clones were developed by selection by G418. Biological characteristics of one of the 6 cell lines which highly expressing TSLC1, namely, the M8T were studied. Cell growth was analyzed with MTT assay. 2 x 10^7cells suspended in 0.2 mL phosphate buffered saline （PBS） were injected into the two flanks of 5-6-week-old female BALB/C nu/nu athymic nude mice. The volumes of subcutaneous of tumor growth were evaluated and calculated by the formula V= Length x Width x Height x 0.5 once a week. Results： The MST cell subline which stably expressing TSLC1 was characterized by Western blot. The genetic stability and purity of M8T cells were stable. TSLC1 significantly suppressed the growth of M8T cells in vitro. Moreover, the tumorigenicity of MST cells was suppressed in vivo. Conclusion： The osteosarcoma cell sublines MST which stably expressing TSLC1 had been successfully established. The ability of growth and metastasis of MST was significantly suppressed both in vitro and in vivo.

TonEBP expression is essential in the IL-1β–induced migration and invasion of human A549 lung cancer cells

Lung cancer has the highest mortality rate among all cancers,in part because it readily metastasizes.The tumor microenvironment,comprising blood vessels,fibroblasts,immune cells,and macrophages[including tumor-associated macrophages(TAMs)],is closely related to cancer cell growth,migration,and invasion.TAMs secrete several cytokines,including interleukin(IL)-1β,which participate in cancer migration and invasion.p21-activated kinase 1(PAK1),an important signaling molecule,induces cell migration and invasion in several carcinomas.Tonicityresponsive enhancer-binding protein(TonEBP)is also known to participate in cancer cell growth,migration,and invasion.However,the mechanisms by which it increases lung cancer migration remain unclear.Therefore,in this study,we aimed to elucidate the mechanisms by which IL-1βand TonEBP affect lung cancer cell migration and invasion.We found that A549 cocultured-MΦ-secreted IL-1βinduced A549 cell migration and invasion via the PAK1 pathway.TonEBP deficiency reduced A549 cell migration and invasion and increased responsiveness to IL-1β–induced migration and invasion.PAK1 phosphorylation,which was promoted by IL-1β,was reduced when TonEBP was depleted.These results suggest that TonEBP plays an important role in IL-1βinduction and invasiveness of A549 cells via the PAK1 pathway.These findings could be valuable in identifying potential targets for lung cancer treatment.

Identification of a potential tumor suppressor gene, UBL3, in non-small cell lung cancer

Objective: Oncogenes have been shown to be drivers of non-small cell lung cancer(NSCLC), yet the tumor suppressing genes involved in lung carcinogenesis remain to be systematically investigated. This study aimed to identify tumor suppressing ubiquitin pathway genes(UPGs) that were critical to lung tumorigenesis.Methods: The 696 UPGs were silenced by an siRNA screening in NSCLC cells;the potential tumor suppressing UPGs were analyzed, and their clinical significance was investigated.Results: We reported that silencing of 11 UPGs resulted in enhanced proliferation of NSCLC cells, and four UPGs(UBL3, TRIM22, UBE2 G2, and MARCH1) were significantly downregulated in tumor samples compared to that in normal lung tissues and their expression levels were positively associated with overall survival(OS) of NSCLC patients. Among these genes, UBL3 was the most significant one. UBL3 expression was decreased in tumor samples compared to that in paired normal lung tissues in 59/86(68.6%) NSCLCs, was correlated with TNM stage and sex of NSCLC patients, and was significantly higher in non-smoking patients than in smoking patients. Silencing UBL3 accelerated cell proliferation and ectopic expression of UBL3 suppressed NSCLC in vitro and in vivo.Conclusions: These results showed that UBL3 represented a tumor suppressor in NSCLC and may have potential for use in therapeutics and for the prediction of clinical outcome of patients.

miR-30a-5p/PHTF2 axis regulates the tumorigenesis and metastasis of lung adenocarcinoma

Background:Lung adenocarcinoma is a very pervasive histological form of lung cancers,and inhibiting metastasis is crucial for effective treatment.In this investigation,we explored the functional interaction of miR-30a-5p and the putative transcription factor 2 of the homeodomain(PHTF2)in dictating the aggressiveness and metastasis of lung adenocarcinoma.Method:We collected clinical samples to evaluate the expression patterns of miR-30a-5p and PHTF2 in lung adenocarcinoma along with normal tissues.Cellular experiments including cell count kit(CCK)-8 growth assay,apoptosis analysis,migration and invasion examinations were performed to assess the aggressiveness of lung adenocarcinoma cells.Furthermore,we examined tumorigenesis and metastasis in a nude mouse model.Results:MiR-30a-5p exhibited downregulation pattern in lung adenocarcinoma samples.Transfection of miR-30a-5p mimic in lung adenocarcinoma cells resulted in the suppression of malignant characteristics.Notably,the administration of miR-30a-5p mimic also curbed tumorigenesis and metastasis of lung adenocarcinoma cells in animal model.Moreover,PHTF2 was found to be a molecular target of miR-30a-5p.Upregulating PHTF2 counteracted the tumor-suppressive effect of the miR-30a-5p mimic.Conclusion:miR-30a-5p functions as a tumor-suppressive molecule while PHTF2 acts as an oncogenic factor in the development and metastasis of lung adenocarcinoma.Therefore,targeting miR-30a-5p and PHTF2 could be developed into a promising therapeutic approach for inhibiting metastasis in lung adenocarcinoma.

Cancer/testis antigen, Kita-Kyushu lung cancer antigen-1 and ABCD stratification for diagnosing gastric cancers

BACKGROUND The ABCD stratification[combination of serum pepsinogen(PG)levels and titers of antibody(immunoglobulin G,IgG)against Helicobacter pylori(H.pylori)]is effective for the classification of individuals at risk of developing gastric cancer(GC).The Kita–Kyushu lung cancer antigen-1(KK-LC-1)is a Cancer/Testis antigen frequently expressed in GC.AIM To evaluate the effectiveness of KK-LC-1 and ABCD stratification in the diagnosis of GC.METHODS We analyzed the gene expression of KK-LC-1 in surgical specimens obtained from GC tumors.The levels of serum PG I/PG II and IgG against H.pylori were measured.According to their serological status,the patients were classified into the four groups of the ABCD stratification.RESULTS Of the 77 examined patients,63(81.8%)expressed KK-LC-1.The IgG titers of H.pylori and PG II were significantly higher in patients expressing KK-LC-1 than those measured in patients not expressing KK-LC-1(P=0.0289 and P=0.0041,respectively).The expression of KK-LC-1 in group C[PG method(+)/H.pylori infection(+)]was as high as 93.9%high.KK-LC-1 was also detected in group A[-/-].CONCLUSION The KK-LC-1 expression in GC was associated with H.pylori infection and atrophic status,so that,KK-LC-1 may be a useful marker for the diagnosis of GC.

Oncoprotein HBXIP promotes tumorigenesis through MAPK/ERK pathway activation in non-small cell lung cancer

Objective:The oncoprotein,hepatitis B X-interacting protein(HBXIP),has been reported to play an important role in human malignancies.However,its functions in non-small cell lung cancer(NSCLC)are poorly understood.The goal of the present study was to identify the role of HBXIP in the regulation of NSCLC development.Methods:The level of HBXIP expression in NSCLC tissue was assessed by immunohistochemical and Western blot analyses,and its relationships with clinicopathological features and outcomes were statistically evaluated.The effects of HBXIP on NSCLC cell progression were assessed through cell viability,colony formation,and flow cytometry analyses in vitro.The mechanism by which HBXIP regulated the MAPK pathway was studied by Western blot,immunofluorescence,and immunoprecipitation assays.In addition,in vivo experiments were performed to evaluate the progression of NSCLC and ERK signaling pathway activation after HBXIP knockdown.Results:HBXIP was overexpressed in human NSCLC and was correlated with the invasiveness of NSCLC.The high expression of HBXIP in NSCLC was significantly correlated with gender(P=0.033),N stage(P=0.002),and tumor-node-metastasis stage(P=0.008).In vitro experiments using an NSCLC cell line revealed that HBXIP knockdown resulted in the suppression of cell proliferation and colony formation,which was consistent with the enhanced cell cycle arrest in G1 phase.The results of a mechanistic investigation suggested that binding of HBXIP to MEK1 protein promoted MAPK/ERK signaling pathway activation in NSCLC by preventing the proteasome-mediated degradation of MEK1.In addition,the results obtained using in vivo subcutaneous tumor xenografts confirmed that HBXIP deficiency decreased MEK1 protein levels and NSCLC tumor growth.Conclusions:Taken together,our results showed that the HBXIP-MEK interaction promoted oncogenesis via the MAPK/ERK pathway,which may serve as a novel therapeutic target for cancers in which MAPK/ERK signaling is a dominant feature.

HYPERMETHYLATION OF p14^(ARF) PROMOTER REGION AND EXPRESION OF p14^(ARF) GENE PRODUCT IN NON-SMALL CELL LUNG CANCER

Objective： This study was designed to investigate promoter methylation status and protein expression of p14^ARF gene in non-small cell lung cancer, and value the role of p14^ARF promoter methylation in carcinogenesis of non-small cell lung cancer. Methods： Promoter methylation status and protein expression of p14^ARF gene in 40 cases of non-small cell lung cancer were analyzed by methylation specific polymerase china reaction （MSP）, restriction enzyme-related polymerase chain reaction （RE-PCR） and immunohistochemistry （IHC）. Results： The positive rates of p14^ARF promoter methylation in tumor tissues and normal tissues adjacent to cancer were 17.5% （7/40） and 2.5% （1/40） respectively. There were statistically significant differences between them, P〈0.05. The results of RE-PCR were consistent with that of MSP. The expression rate of p14^ARF protein in tumor tissues was significantly lower than that in normal tissues adjacent to cancer, p〈0.01. Promoter methylation status and protein expression of p14^ARF gene in non-small cell lung cancer showed significantly an inverse correlation （r=-0.56, P〈0.01）, and both of them did not relate statistically with the clinicopathologic characteristics of patients such as histological classification, clinical stage, differentiation grade and lymph node involvement. Conclusion： Promoter methylation is a crucial mechanism of inactivation of p14^ARF gene. Promoter methylation of p14^ARF gene might he involved in carcinogenesis of non-small cell lung cancer, and is an early event in development process of non-small cell lung cancer. It might be used as a new target in gene treatments in the future.

Influence on radiosensitivity of lung glandular cancer cells when ERCC1 gene silenced by targeted siRNA

Objective:To identify the influence on radiosensitivity of lung glandular cancer cells when excisions repair cross-complementing group1(ERCC1) gene was silenced by targeted siR NA.Methods:siR NA which targeting to ERCC1 and control siR NA was designed and synthesized.The human lung glandular cancer SPC-A-1 cells was transfected.A total of 56 nude mice were divided into two groups,and two kinds of SPC-A-1 cells were transplanted to armpit of right forelimb,to establish the nude mice subcutaneous xenotransplanted tumor model of human lung glandular cancer cells.After the tumor was developed,the nude mice were randomly divided into four groups and accepted different doses of X-Ray radiation,then the change of tumor volume,survival time of mice in every group were recorded and the average lifetime was calculated.Twenty-one days later of X-ray experiment,two mice were taken and sacrificed in each group and the tumors organizations were stripped.The cell apoptosis rate and cell cycle distributions were obtained by FCM(flow cytometry).Results:The volume of tumor which ERCC1 gene was silenced was less than single irradiation group after X-ray irradiation,and the growth speed was slower and the lifetime of mice was lengthened as well(P<0.05).The cells apoptosis rate and the rate of G2/M cells which ERCC1 gene was silenced were higher than the same dose control group and the rate of G_1 cells were lower,which indicated that the cells could be stopped at G_2/M point,the cell proliferation was inhibited,the cell apoptosis was promoted and the radiation sensitivity was improved after the ERCC1 was silenced.Conclusions:The radiation sensitivity of lung glandular tumor could be improved after the ERCC1 gene was silenced by siR NA.

Effects of multiple tumor suppressor 1 on the proliferation of human ovarian cancer HO-8910 cells

objective: To investigate the effects of multiple tumor suppressor 1 on the proliferation of ovarian cancer cell lines. Methods: Growth characteristics of HO-8910 (without p16 gene expression ), 8910-p16 (trans fected with p16 gene) and 8910-pcDNA3 (transfected with the vector pcDNA3) cells were studied by comparison of the cell growth curves. DNA synthesis was also compared among the 3 kinds of cells. Results: After trans fected with p16 gene, the 8910-p16 cells were markedly inhibited in both the proliferation and DNA synthesis. There was no significant difference between the 8910-pcDNA3 cells and the HO-8910 cells. Conclusion: Multiple tumor suppressor 1 can inhibit the proliferation and DNA synthesis of human ovarian cancer HO-8910 cells.

Biallelic Inactivation of hMLH1 by Hypermethylation and Loss of Heterozygosity in Non-Small Cell Lung Cancer

OBJECTIVE To investigate the mechanism of hMLH1 deregulation in non-small cell lung cancer （NSCLC）. METHODS A genetic and epigenetic study of the hMLH1 gene was performed using surgical primary tumors from 40 NSCLC patients and their corresponding noncancerous tissues. The molecular alterations examined included promoter methylation by Hpa Ⅱ/Msp Ⅰ- based PCR analysis, loss of heterozygosity （LOH） by D3S1621 locus PCR-electrophoresis-silver staining, as well as the loss of protein expression by immunohistochemical analysis. RESULTS The frequencies of hypermethylation, LOH and loss of protein expression of hMLH1 were 67.5% （27/40）, 65% （26/40） and 72.5% （29/ 40）, respectively. Among 26 hMLH1 gene LOH （＋） cases, 21 （80.8%） showed hypermethylation, which was significantly higher than the group of LOH （-）. The frequency of the double inactivation of the hMLH1 gene by hypermethylation and LOH related to a loss of protein expression of 72.4% （21/29）. CONCLUSION Biallelic inactivation of the hMLH1 gene by hypermethylation and LOH most likely will cause loss of hMLH1 protein expression and play an important role in the development of NSCLC. Therefore, controlling and monitoring for hypermethylation of the hMLH1 gene may be partially useful for treatment and early diagnosis of NSCLC.

Effects of Yiqi Yangyin Prescription combined with radiotherapy on the tumor load and anti-tumor immune response of patients with advanced lung cancer

Objective: To investigate the effects of Yiqi Yangyin Prescription combined with radiotherapy on the tumor load and anti-tumor immune response of patients with advanced lung cancer. Methods: A total of 78 patients with advanced non-small cell lung cancer who were diagnosed and treated in our hospital between August 2015 and January 2017 were reviewed and then divided into the routine group (n=41) who received chemoradiotherapy alone and the Yiqi Yangyin Prescription group (n=37) who received chemoradiotherapy combined with Yiqi Yangyin Prescription therapy. The differences in serum levels of tumor markers, Th1/Th2 cytokines and Th17/Treg cytokines were compared between the two groups before and after treatment. Results: After treatment, serum tumor markers CA15-3, Cyfra21-1, HSP90 and CEA levels of Yiqi Yangyin Prescription group were lower than those of routine group;serum Th1 cytokines IFN-γ and IL-2 levels were higher than those of routine group while Th2 cytokines IL-4 and IL-5 levels were lower than those of routine group;serum Th17 cytokines IL-17 and IL-23 as well as Treg cytokines IL-10 and TGF-β levels were lower than those of routine group. Conclusion: Routine chemoradiotherapy combined with Yiqi Yangyin Prescription therapy can effectively reduce the tumor load and optimize the anti-tumor immune function in patients with advanced non-small cell lung cancer.

Effects of eEF1A1 expression on cancer cell load and proliferation activity in lung cancer lesions

Objective: To investigate the effects of eEF1A1 expression on cancer cell load and proliferation activity in lung cancer lesions. Methods: A total of 200 patients with non-small cell lung cancer who received radical operation for lung cancer in our hospital were enrolled in lung cancer group, and 78 patients with pulmonary bullae who accepted emergency surgical treatment in our hospital during the same period were enrolled in pulmonary bullae group. Differences in eEF1A1 and proliferation gene expression in lesion tissue were compared between the two groups, and the contents of serum tumor markers and angiogenesis indexes were detected. Pearson test was used to evaluate the correlation of eEF1A1 expression with tumor markers, angiogenesis indexes and proliferation genes in the lesions of patients with non-small cell lung cancer. Results: eEF1A1 expression in lesion tissue of lung cancer group was higher than that of pulmonary bullae group;serum CYFRA21-1, ProGRP, CEA, SCC Ag, HIF-1 and VEGF levels of lung cancer group were higher than those of pulmonary bullae group and positively correlated with the eEF1A1 expression;LRRC3B, TUSC3 and VPS33B mRNA expression in lesion tissue of lung cancer group were lower than those of pulmonary bullae group and negatively correlated with the eEF1A1 expression whereas MACC1 and RACK1 mRNA expression were higher than those of pulmonary bullae group and positively correlated with the eEF1A1 expression. Conclusion: eEF1A1 is highly expressed in non-small cell lung cancer tissues, and the specific expression can objectively reflect the tumor load and the proliferation activity of tumor cells.

Underexpression of LATS1 TSG in colorectal cancer is associated with promoter hypermethylation

AIM:To investigate large tumor suppressor 1 (LATS1 ) expression, promoter hypermethylation, and microsatellite instability in colorectal cancer (CRC).METHODS:RNA was isolated from tumor tissue of 142 CRC patients and 40 colon mucosal biopsies of healthy controls. After reverse transcription, quantitative polymerase chain reaction (PCR) was performed, and LATS1 expression was normalized to expression of the ACTB and RPL32 housekeeping genes. To analyze hypermethylation, genomic DNA was isolated from 44 tumor CRC biopsies, and methylation-specific PCR was performed. Microsatellite instability (MSI) status was checked with PCR using BAT26, BAT25, and BAT40 markers in the genomic DNA of 84 CRC patients, followed by denaturing gel electrophoresis. RESULTS:Decreased LATS1 expression was found in 127/142 (89.4%) CRC cases with the average ratio of the LATS1 level 10.33 ± 32.64 in CRC patients vs 32.85 ± 33.56 in healthy controls. The lowest expression was found in Dukes' B stage tumors and G1 (welldifferentiated) cells. Hypermethylation of the LATS1 promoter was present in 25/44 (57%) CRC cases analyzed. LATS1 promoter hypermethylation was strongly associated with decreased gene expression; methylated cases showed 162× lower expression of LATS1 than unmethylated cases. Although high-grade MSI (mutation in all three markers) was found in 14/84 (17%) cases and low-grade MSI (mutation in 1-2 markers) was found in 30/84 (36%) cases, we found no association with LATS1 expression. CONCLUSION:Decreased expression of LATS1 in CRC was associated with promoter hypermethylation, but not MSI status. Such reduced expression may promote progression of CRC.

Tumor-related cytokine release syndrome in a treatment-naïve patient with lung adenocarcinoma:A case report

BACKGROUND Cytokine release syndrome(CRS)is defined as systemic inflammation that usually occurs following chimeric antigen receptor T-cell therapy administration;however,it has not been reported in patients with untreated non-small cell lung cancer to date.CASE SUMMARY A 44-year-old nonsmoking woman presented to the hospital due to fever,palpitation,nausea,and cough for 1 mo and was diagnosed with stage cT3N3M0(IIIc)adenocarcinoma of the lung.Auxiliary examinations revealed elevated cytokine[tumor necrosis factor-α,interleukin(IL)-1β,and IL-6]and inflammatory factor levels,which decreased after treatment with corticosteroids and immunoglobulin and when tumor growth was controlled following chemotherapy,radiotherapy,and antiangiogenesis therapy.However,tumor recurrence was observed.After administration of nivolumab as third-line treatment,the patient’s condition was transiently controlled;however,CRS-like symptoms suddenly emerged,which led to a resurgence of cytokines and inflammatory factors and rapid death.CONCLUSION CRS can develop in treatment-naïve lung cancer patients.Patients with tumorrelated CRS may be at risk of CRS recurrence,aggravation,and onset of immune checkpoint inhibitor-related adverse events.

Re-expression of Cell Adhesion Molecule Inhibits Growth and Induces Apoptosis of Human Pancreatic Cancer Cell Line PANC-1

This study examined the expression of cell adhesion molecule 1 （CADM1） in pancreatic cancer and the possible mechanism. The expression of CADM 1 was detected by immunohistochemistry in tissues of pancreatic cancer, pancreatitis, and normal pancreas. The plasmid pcDNA3.1-Hy- gro（＋）/CADM1 was transfected into PANC-1 cells （a pancreatic cancer cell line）. The expression of CADM1 in the transfected cells was determined by RT-PCR and Western blotting. Cell growth was measured by the MTT method and cell apoptosis by flow cytometry. The results showed that CADM1 was weakly expressed in tissues of pancreatic cancer in contrast to its high expression in normal pancreatic and pancreatitis tissues. The expression level of CADM in pancreatic caner was intensely correlated with the differentiation degree, lymph node metastasis and TNM stages. The growth of CADMl-transfected PANC-1 cells was significantly suppressed in vitro by a G1 cell cycle arrest and apoptosis occurrence. It was concluded that re-expression of CADM1 inhibits the growth of pancreatic cancer cells and induces their apoptosis in vitro. As a tumor suppressor gene, CADM1 plays an important role in the occurrence, progression and metastasis of pancreatic cancer.

Down-Regulation of GKN1 in Gastric Cancer Is Not Associated with the RUNX3 Expression

Gastrokine 1 (GKN1) is a gastric mucosal protein highly expressed and secreted in normal individuals but during Helicobacter pylori infection or in gastric carcinogenesis it is strongly down-regulated or totally absent. In gastric cancer, the GKN1 gene is silenced through an epigenetic mechanism most likely mediated by a transcription factor that promotes on GKN1 promoter the activity of the enzymes SUV39H1 and HDACs. Because RUNX3 is a potential candidate in the regulation of molecular carcinogenesis process of stomach cancers, we tried to assess if RUNX3 could be involved in GKN1 down-regulation in GC. 17 paired of non-tumoral and tumoral surgical specimens from patients with gastric cancer were analyzed for GKN1 and RUNX3 by Western blotting and chromatin immunoprecipitation (Chip) assays. The overall results indicated that RUNX3 expression was not associated with the down-regulation of GKN1. The expression levels of RUNX3 in non-tumoral and tumoral samples suggest that RUNX3 does not act as a tumor suppressor but that it might play a complex oncogenic role in gastric cancer cells.

非小细胞肺癌中p14^(ARF)与p16^(INK4a)和p53蛋白表达及其相互关系的研究

目的:检测p14ARF、p16INK4a和p53蛋白在非小细胞肺癌中表达情况,对它们在非小细胞肺癌发生过程中的作用及其相互关系进行初步探讨。方法:应用免疫组化方法检测了40例非小细胞肺癌组织标本的p14ARF、p16INK4a和p53基因的表达水平。结果:非小细胞肺癌组织中p14ARF、p16INK4a和p53蛋白总阳性率分别是72.5%,50.0%和52.5%,与癌旁正常组织(分别是95.0%,92.5%,2.5%)有显著性差异(P<0.01);非小细胞肺癌中p14ARF和p53蛋白表达异常与肿瘤分期、组织类型、分化程度、淋巴结转移情况无显著相关性;p16INK4a蛋白表达水平与组织分化程度相关(P<0.05),但与肿瘤分期、组织类型、淋巴结转移情况无关;p14ARF和p53蛋白在组织中表达呈负相关,r2=-0.475(P<0.01),而p14ARF和p16INK4a蛋白表达无关;p14ARF蛋白在Ⅰ期病例中强阳性(+++)占阳性比例为41.7%,在Ⅲ期中强阳性(+++)比例为10.0%。结论:p14ARF、p16INK4a和p53三个基因在NSCLC发生过程中起重要作用,特别在肿瘤早期可能起到更为重要的作用;p14ARF和p53基因在同一调节通路上;p14ARF、p16INK4a和p53蛋白可以作为非小细胞肺癌早期生物学检测指标。

抑癌基因 ING1在大肠癌中的表达及预后意义

目的：探讨ING1基因在散发性大肠癌中的表达与预后及多个临床病理变量之间的关系，并分析其在大肠癌预后的危险因素中是否具有显著意义。方法应用定量RT－PCR方法检测82例大肠癌手术切除标本及相应的癌旁组织中ING1 mRNA的表达水平，分析其表达的差异，研究其表达水平与大肠癌患者临床病理特征及预后的关系。结果（1）ING1 mRNA在大肠癌及癌旁组织中均可被检出，同一配对组织相比，癌旁组织中ING1表达量明显高于肿瘤原发灶；（2） ING1 mRNA的表达与大肠癌的浸润层次、淋巴结转移、远处转移以及TNM分期密切相关；（3）肿瘤组织与癌旁组织ING1表达量相比，比值越低其DFS也越低（P＜0．0001）；（4）经单因素及多因素COX模型分析后显示，ING1作为候选抑癌基因可作为大肠癌预后的独立预测因素（ P＜0．0001）。结论 ING1的过度表达是大肠癌发生过程中的分子事件，可能参与大肠癌的发展过程。 ING1可作为判断大肠癌预后的重要分子标志。

抑癌基因beclin1在子宫内膜异位症及非子宫内膜异位症患者中差异表达的意义

目的研究子宫内膜异位症(cndometriosis,Ems)组织中抑癌基因beclin 1的mRNA和蛋白表达情况。方法利用RT-PCR和Western Blot免疫印迹的方法,对EMs的异位内膜组织、在位内膜组织和非EMs在位内膜组织(正常内膜组织)进行mRNA和蛋白表达的半定量分析。利用原位杂交和免疫组化技术对研究各组进行定性及定位分析。结果①beclin 1 mRNA和蛋白在各组中均有表达,异位内膜的表达高于在位内膜和正常内膜,差异有显著性(P<0.005),在位内膜组与正常对照组比较,beclin 1的表达无明显差异(P>0.05);②原位杂交阳性率分别为非EMs组95.1%,EMs异位内膜组100%,EMs在位内膜组91.4%;③免疫组化阳性率分别为非EMs组91.7%,EMs异位内膜组100%,EMs在位内膜组93.4%;④beclin 1的DNA与蛋白表达一致;⑤7例恶变的异位内膜beclin 1免疫组化检测,5例呈阴性,2例呈弱阳性表达。结论抑癌基因beclin 1在子宫内膜组织中均表达,且在异位内膜中呈表达上调,而在恶变的异位内膜中多呈阴性表达,提示beclin 1基因可能与子宫内膜异位症不孕及恶变相关。

肺癌人源性肿瘤组织异种移植模型的组织学变化及其p63、napsinA和TTF-1的表达差异

目的建立肺癌人源性肿瘤组织异种移植(patient-derived tumor xenograft,PDX)裸鼠模型,检测原代肺癌组织标本和第三代(F3)PDX模型组织标本内组织形态结构的变化和p63、napsin A和TTF-1的表达差异性。方法取临床肺癌新鲜手术切除标本12例,裸鼠皮下移植建立PDX模型,HE染色比较原代患者肿瘤组织与移植瘤组织结构和细胞形态变化与否;免疫组化的方法检测原代组织和F3代组织标本内p63、napsin A和TTF-1的表达水平的差异性。显微镜观察。结果成功建立传至第三代的PDX模型5例,4例肺鳞癌,1例肺腺癌。PDX模型和患者原代肿瘤具有相同的组织学特点和排列结构;p63在肺鳞癌患者及其PDX模型癌细胞中表达阳性率为84.3%和96%,两者之间差异无显著性(P>0.05),而在肺腺癌中呈阴性表达。Napsin A在肺腺癌患者及其PDX F3癌细胞中阳性表达率分别为66%和72.4%,两者之间差异无显著性(P>0.05)。TTF-1在肺腺癌患者及其PDX F3癌细胞中阳性表达率分别为91%和85%,两者之间差异也无显著性(P>0.05)。Napsin A和TTF-1在肺鳞癌中均呈阴性表达。结论本实验室所建立的肺癌PDX模型基本保留了部分原代肿瘤的组织学特性,为临床前药效的个性化筛选评估以及生物标志物的鉴定提供了有效的研发资源。