Treatment and surveillance for non-muscle-invasive bladder cancer:a clinical practice guideline(2021 edition)

Non-muscle invasive bladder cancer(NMIBC)is a major type of bladder cancer with a high incidence worldwide,resulting in a great disease burden.Treatment and surveillance are the most important part of NIMBC management.In 2018,we issued“Treatment and surveillance for non-muscle-invasive bladder cancer in China:an evidencebased clinical practice guideline”.Since then,various studies on the treatment and surveillance of NMIBC have been published.There is a need to incorporate these materials and also to take into account the relatively limited medical resources in primary medical institutions in China.Developing a version of guideline which takes these two issues into account to promote the management of NMIBC is therefore indicated.We formed a working group of clinical experts and methodologists.Through questionnaire investigation of clinicians including primary medical institutions,24 clinically concerned issues,involving transurethral resection of bladder tumor(TURBT),intravesical chemotherapy and intravesical immunotherapy of NMIBC,and follow-up and surveillance of the NMIBC patients,were determined for this guideline.Researches and recommendations on the management of NMIBC in databases,guideline development professional societies and monographs were referred to,and the European Association of Urology was used to assess the certainty of generated recommendations.Finally,we issued 29 statements,among which 22 were strong recommendations,and 7 were weak recommendations.These recommendations cover the topics of TURBT,postoperative chemotherapy after TURBT,Bacillus Calmette–Guérin(BCG)immunotherapy after TURBT,combination treatment of BCG and chemotherapy after TURBT,treatment of carcinoma in situ,radical cystectomy,treatment of NMIBC recurrence,and follow-up and surveillance.We hope these recommendations can help promote the treatment and surveillance of NMIBC in China,especially for the primary medical institutions.

Role of endoscopic ultrasound for pre-intervention evaluation in early esophageal cancer

BACKGROUND Endoscopic ultrasound(EUS)stands as an accurate imaging modality for esophageal cancer staging,however utilization of EUS in early-stage cancer management remains controversial.Identification of non-applicability of endoscopic interventions with deep muscular invasion with EUS in pre-intervention evaluation of early-stage esophageal cancer is compared to endoscopic and histologic indicators.AIM To display the role of EUS in pre-intervention early esophageal cancer staging and how the index endoscopic features of invasive esophageal malignancy compare for prediction of depth of invasion and cancer management.METHODS This was a retrospective study of patients who underwent pre-resection EUS after a diagnosis of esophageal cancer at a tertiary medical center from 2012 to 2022.Patient clinical data,initial esophagogastroduodenoscopy/biopsy,EUS,and final resection pathology reports were abstracted,and statistical analysis was conducted to assess the role of EUS in management decisions.RESULTS Forty nine patients were identified for this study.EUS T stage was concordant with histological T stage in 75.5%of patients.In determining submucosal involvement(T1a vs T1b),EUS had a specificity of 85.0%,sensitivity of 53.9%,and accuracy of 72.7%.Endoscopic features of tumor size>2 cm and the presence of esophageal ulceration were significantly associated with deep invasion of cancer on histology.EUS affected management from endoscopic mucosal resection/submucosal dissection to esophagectomy in 23.5%of patients without esophageal ulceration and 6.9%of patients with tumor size<2 cm.In patients without both endoscopic findings,EUS identified deeper cancer and changed management in 4.8%(1/20)of cases.CONCLUSION EUS was reasonably specific in ruling out submucosal invasion but had relatively poor sensitivity.Data validated endoscopic indicators suggested superficial cancers in the group with a tumor size<2 cm and the lack of esophageal ulceration.In patients with these findings,EUS rarely identified a deep cancer that warranted a change in management.

Intravesical bacillus Calmette-Guerin(BCG)in treating non-muscle invasive bladder cancer—analysis of adverse effects and effectiveness of two strains of BCG(Danish 1331 and Moscow-I)

Objective:To compare the differences in adverse effects and efficacy profile between bacillus Calmette-Guerin(BCG)Danish 1331 and BCG Moscow-I strain in management of non-muscle invasive bladder cancer.Methods:Clinical data of 188 cases of non-muscle invasive bladder cancer treated with BCG between January 2008 and December 2018 in our institute were collected prospectively and analysed retrospectively,and 114 patients who completed a minimum of 12 months of follow-up were analysed.Patient and tumor characteristics,strain of BCG,adverse effects,and tumor progression were included for analysis.Intravesical BCG was instilled in intermediate-and high-risk patients.Six weeks of induction BCG,followed by three weekly maintenance BCG at 3,6,12,18,and 24 months was advised in high-risk patients.Results:Overall 68 patients received BCG Danish 1331 strain and 46 patients received Moscow-I strain.Patient and tumor characteristics were well balanced between the two groups.The median follow-up period was 42.5 months and 34.5 months in Danish 1331 and Moscow-I groups,respectively.Adverse events like dropout rate,antitubercular treatment requirement,and need of cystectomy were higher in Moscow-I group(n=31,67.4%)when compared to Danish 1331 strain(n=33,48.5%)(p=0.046).On direct comparison between Danish 1331 and Moscow-I strain,there was similar 3-year recurrence-free survival(80.0%vs.72.9%)and 3-year progression-free survival(96.5%vs.97.8%).Conclusion:Study results suggest no significant differences between Danish 1331 and Moscow-I strain in recurrence-free survival and progression-free survival,but a significantly higher incidence of moderate to severe adverse events in BCG Moscow-I strain.

Role of gemcitabine and cisplatin as neoadjuvant chemotherapy in muscle invasive bladder cancer: Experience over the last decade

Objective:Neoadjuvant chemotherapy followed by radical cystectomy is considered the standard of care for patients with muscle invasive bladder cancer.In the last decade,interest in neoadjuvant chemotherapy has slowly shifted from methotrexate,vinblastine,doxorubicin and cisplatin regime to gemcitabine and cisplatin regime.There are many publications on gemcitabine and cisplatin regime in literature which cover different aspects of treatment.This review aims to summarise the findings published so far on gemcitabine and cisplatin regime and present it in a concise manner.Methods:A systematic literature review was conducted searching the PubMed
database in December 2016 using the medical subject heading(MeSH)with the terms gemcitabine,cisplatin,chemotherapy,muscle invasive bladder cancer,and neoadjuvant.All relevant studies were included and results were analysed.Results:A total of 13 studies were included which published between 2007 and 2015.These 13 studies comprised of 754 subjects suffering from muscle invasive bladder cancer.The proportion of male patients ranged from 60%to 86.4%and the median age ranged from 54.2 to 77.3 years in various studies.Complete pathological response(pT0)was seen in 30.0%of patients and pathological downstaging(<pT2)was seen in 48.67%of patients.Conclusion:As per latest guidelines,neoadjuvant chemotherapy is recommended for patients with muscle invasive bladder cancer.There is substantial pathological downstaging with low toxicity in patients of muscle invasive bladder cancer who receive neoadjuvant gemcitabine and cisplatin regime.

Molecular mechanisms and novel therapeutic strategies of BCG-unresponsive non-muscle invasive bladder cancer: Emerging immunotherapy has become a new choice?

Objective:THigh-risk non-invasive bladder cancer(NMIBC)has a high rate of recurrence and disease progression.At present,there are still insufficient effective prevention and treatment methods,especially for patients who have failed BCG treatment.This article reviews the research progress of the molecular mechanism of BCG unresponsive NMIBC,and summarizes the current status and prospects of emerging therapeutic strategies represented by immunotherapy,providing a theoretical basis for the immunotherapy of BCG non-reactive NMIBC.Methods:We searched the PubMed and CNKI journal full-text database search system for keywords"non-muscle invasive bladder cancer,BCG unresponsive,disease recurrence,disease progression,and immunotherapy"with 126 English and 538 Chinese articles.The literature,as well as the relevant clinical research in ClinicalTrials.gov,were integrated together to obtain the results.Results:Immunotherapy was performed in various types of tumors,and the use of immunotherapeutic drugs with different oncotargets administered alone,sequentially or in combination for the treatment of BCG-unresponsive NMIBC have achieved favorable effects,and more Clinical research is still ongoing.Conclusion:Immunotherapy is currently the most promising treatment for cancer,and it is indispensable for patients with NMIBC,both biologically and clinically.We look forward to more laboratory and clinical research in immunotherapy in the future.

Did the Scientific Innovations in the Management of Non-Muscle Invasive Bladder Cancer Patients Improve the Outcome during the Last 2 Decades?

Objectives: Previous reviews reported the outcome of each scientific modality in the management of T1 high-grade bladder cancer. The objective of this review is to assess and evaluate the available scientific modalities used during the last two decades and determine whether they were able to improve the clinical outcome. Literature Search Methodology: A systematic literature review was conducted from 2000-2020 using PubMed, Medline, Embase, and other database sites looking at randomized controlled trials (RCTs), clinical trials, research, review articles, and original articles addressing the different scientific modalities used to diagnose and manage patients with non-muscle invasive Bladder cancer (NMIBC)during the last 2 decades. More than 573 studies were retrieved following the preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines and PICOS criteria (Population, Intervention, Comparators, Outcomes, and Study design). Only 85 articles were selected for review including 19 prospective trials, 44 RCTs, original articles, research articles, one review article, and clinical trials—Retrospective studies were excluded to limit bias as much as possible in the analysis. Results: Randomized controlled trials (RCTs) have become the gold standard for evaluating the efficacy of new treatments. They are considered the highest standard of evidence-based medicine and are the method of choice. Overall, we selected 85 studies for review, among them 63 prospective trials and RCTs, with a total of 21,895 patients, published between 2000 and 2020. Previously conducted studies have shown that identifying rare histological types with poor prognoses can help improve outcomes, mainly the plasmacytoid type. Many articles addressed the role of biomarkers in the early identification of patients with NMIBC for recurrence and progression—P-cadherin expression and others were used to predict recurrence and/or progression with promising results. Despite the need for modifications, risk stratification is an important tool that should be used to improve the outcome of patients with NMIBC. Some found that fluorescence diagnostic cystoscopy (FDC) and Photodynamic diagnosis (PDD) improved recurrence-free survival but not progression and outcome. All authors agree that intravesical BCG is the most effective therapy that changes the course of high-grade T1 mainly progression. Re-TURBT has become one of the recommendations of international societies, but its potential effect on survival improvement is debatable. Most of the articles showed the advantages of early cystectomy in NMIBC but all agree that the selection criteria must be clearly defined. Conclusions: This review analyzed the outcomes provided by the scientific advances in the field of management of NMIBC patients in the last two decades. Patients with T1 bladder cancer have variable outcomes because of tumor heterogeneity and clinical staging. Despite the great development in the field of diagnosis, risk stratification, and management, further large studies are mostly needed to better elucidate this subset of patients and avoid over and under-treatment.

A Th2-score in the tumor microenvironment as a predictive biomarker of response to Bacillus Calmette Guérin in patients with non-muscle invasive bladder carcinoma:A retrospective study

Intravesical Bacillus Calmette Guerin(BCG)is the gold standard therapy for intermediate/high-risk nonmuscle invasive bladder cancer(NMIBC).However,the response rate is~60%,and 50%of non-responders will progress to muscle-invasive disease.BCG induces massive local infiltration of inflammatory cells(Th1)and ultimately cytotoxic tumor elimination.We searched for predictive biomarker of BCG response by analyzing tumor-infiltrating lymphocyte(TIL)polarization in the tumor microenvironment(TME)in pre-treatment biopsies.Pre-treatment biopsies from patients with NMIBC who received adequate intravesical instillation of BCG(n=32)were evaluated retrospectively by immunohistochemistry.TME polarization was assessed by quantifying the T-Bet+(Th1)and GATA-3+(Th2)lymphocyte ratio(G/T),and the density and degranulation of EPX+eosinophils.In addition,PD-1/PD-L1 staining was quantified.The results correlated with BCG response.In most non-responders,Th1/Th2 markers were compared in pre-and post-BCG biopsies.ORR was 65.6%in the study population.BCG responders had a higher G/T ratio and a greater number of degranulated EPX+cells.Variables combined into a Th2-score showed a significant association with higher scores in responders(p=0.027).A Th2-score cut-off value>48.1 allowed discrimination of responders with 91%sensitivity but lower specificity.Relapse-free survival was significantly associated with the Th2-score(p=0.007).In post-BCG biopsies from recurring patients,TILs increased Th2-polarization,probably reflecting BCG failure to induce a pro-inflammatory status and,thus,a lack of response.PD-L1/PD-1 expression was not associated with the response to BCG.Our results support the hypothesis that a preexisting Th2-polarized TME predicts a better response to BCG,assuming a reversion to Th1 polarization and antitumor activity.

Anal Invasive Squamous Cell Cancer and Human Papillomavirus Distribution in HIV-Infected and Non-HIV-Infected Individuals

Invasive-squamous-cell-cancer (ISCC) of the anal canal is an uncommon disease. Human papillomavirus (HPV) is the etiological agent of most of types of ISCC. The incidence of ISCC has been increasing in HIV-infected individuals, even after the introduction of highly active antiretroviral therapy. The aim of this study was to analyze biopsy specimens from patients diagnosed with ISCC at a tertiary hospital from 1983 to 2012 in order to detect HPV-DNA. Methods: Formaldehyde-fixed, paraffin-embedded specimens from patients with ISCC underwent HPV-DNA genotyping using multiplex PCR assay. Results: A total of 31 cases were collected;10 were HIV-infected (9 men, 1 woman) and 21 non-HIV-infected (11 men, 10 women). HPV infection was detected in 87.5% (7/8) of the HIV-infected patients (DNA from 2 biopsies was degraded) and 76.2% (16/21) of non-HIV-infected individuals. Multiple-type infections were only found in 28.6% (2/7) of the HIV-infected patients (no multiple-type infections in non-HIV-infected individuals). The most prevalent type was HPV-16: 50% (4/8) in the HIV-infected group (57% [4/7] of the HPV-positive samples) and 66.7% (14/21) in the non-HIV-infected group (87.5% (14/16) of the HPV-positive samples). Remarkably, 37.5% (3/8) of the HIV-infected group had high-risk HPV types not included in the vaccines (HPV-33, 51, 52, and 66) compared with 4.8% in the non-HIV-infected group (1/21, HPV-52). All cases of anal ISCC in HIV-infected patients were recorded in the highly active antiretroviral therapy era. Conclusion: HIV-infected patients presented anal ISCC with a higher proportion of high-risk HPV types not covered by the conventional vaccines than non-HIV-infected individuals.

Minimally invasive esophagectomy for the treatment of esophageal cancer:a report of 81 cases

Objective To assess the feasibility and clinical efficacy of minimally invasive esophagectomy for esophageal cancer. Methods From July 2007 to December 2009,eighty-one patients with esophageal cancer received combined thoracoscopic and laparoscopic esophagectomy with anastomosis in the neck. All clinical data were retrospectively reviewed. Results The median operative

Individualized prediction of perineural invasion in colorectal cancer: development and validation of a radiomics prediction model

Objective： To develop and validate a radiomics prediction model for individualized prediction of perineural invasion（PNI） in colorectal cancer（CRC）.Methods： After computed tomography（CT） radiomics features extraction, a radiomics signature was constructed in derivation cohort（346 CRC patients）. A prediction model was developed to integrate the radiomics signature and clinical candidate predictors [age, sex, tumor location, and carcinoembryonic antigen（CEA） level]. Apparent prediction performance was assessed. After internal validation, independent temporal validation（separate from the cohort used to build the model） was then conducted in 217 CRC patients. The final model was converted to an easy-to-use nomogram.Results： The developed radiomics nomogram that integrated the radiomics signature and CEA level showed good calibration and discrimination performance [Harrell＇s concordance index（c-index）： 0.817; 95% confidence interval（95% CI）： 0.811–0.823]. Application of the nomogram in validation cohort gave a comparable calibration and discrimination（c-index： 0.803; 95% CI： 0.794–0.812）.Conclusions： Integrating the radiomics signature and CEA level into a radiomics prediction model enables easy and effective risk assessment of PNI in CRC. This stratification of patients according to their PNI status may provide a basis for individualized auxiliary treatment.

Narrow band imaging for bladder cancer

Narrow band imaging(NBI)is a newly developed technology aiming to provide additional endoscopic information for patients with bladder cancer.This review focuses on the diagnostic accuracy and treatment outcome using NBI cystoscopy for the treatment of nonmuscle invasive bladder cancer.Current results showed improved sensitivity of NBI cystoscopy compared to conventional white light cystoscopy,although lower specificity and increased false-positive results were reported using NBI cystoscopy.The treatment outcome using NBI technology in transurethral resection of bladder tumor had a positive impact while decreased number of residual tumors and tumor recurrence at follow-up were reported.In the future,the application of NBI technology might refine the treatment and follow-up protocol in patients with non-muscle invasive bladder cancer.However,this large scale prospective studies are required to confirm the real cost-effectiveness of this new technology.

Human epidermal growth factor receptor 2 positive rates in invasive lobular breast carcinoma: The Singapore experience

BACKGROUND Invasive lobular carcinomas(ILC)form 5%-10%of breast cancer and rarely show overexpression of human epidermal growth factor receptor 2(HER2).AIM To describe the prevalence and prognostic factors of HER2 positive(HER2+)ILC in an Asian population.METHODS A retrospective review of patients with ILC seen between January 1985 and March 2018 at various SingHealth medical institutions was conducted.Demographic and clinical data were collected from medical records.We examined clinicopathological characteristics and survival in relation to HER2 status.RESULTS A total of 864 patients were included.Prevalence of HER2 positivity was 10.1%(87 patients).Compared with HER2 negative(HER2-)ILC,HER2+ILC was associated with a higher proportion of estrogen receptor negative(24.4%vs 5.9%,P<0.001),progesterone receptor negative(PR-)(40.2%vs 24%,P=0.002)and grade 3 tumours(Grade 3,29.0%vs 10.2%,P<0.001).Overall survival rate was poorer in patients with HER2+compared to HER2-ILC(56.7%vs 72.9%alive at 10 years;hazard ratio 1.87,95%confidence interval:1.21-2.90,P=0.004).Based on multivariate analysis,negative prognostic factors for overall survival included HER2 positivity,PR negativity,older age,Indian ethnicity and higher tumour stage.CONCLUSION Prevalence of HER2+ILC was 10.1%.HER2+ILC was more likely to have poorer prognostic features such as estrogen receptor negative,PR-and higher tumour grade,and have a poorer survival.

A 3D biophysical model for cancer spheroid cell-enhanced invasion in collagen-oriented fiber microenvironment

The process of in situ tumors developing into malignant tumors and exhibiting invasive behavior is extremely complicated.From a biophysical point of view,it is a phase change process affected by many factors,including cell-to-cell,cell-to-chemical material,cell-to-environment interaction,etc.In this study,we constructed spheroids based on green fluorescence metastatic breast cancer cells MDA-MB-231 to simulate malignant tumors in vitro,while constructed a three-dimensional(3D)biochip to simulate a micro-environment for the growth and invasion of spheroids.In the experiment,the 3D spheroid was implanted into the chip,and the oriented collagen fibers controlled by collagen concentration and injection rate could guide the MDA-MB-231 cells in the spheroid to undergo directional invasion.The experiment showed that the oriented fibers greatly accelerated the invasion speed of MDA-MB-231 cells compared with the traditional uniform tumor micro-environment,namely obvious invasive branches appeared on the spheroids within 24 hours.In order to analyze this interesting phenomenon,we have developed a quantitative analyzing approach to explore strong angle correlation between the orientation of collagen fibers and invasive direction of cancer cell.The results showed that the oriented collagen fibers produced by the chip can greatly stimulate the invasion potential of cancer cells.This biochip is not only conducive to modeling cancer cell metastasis and studying cell invasion mechanisms,but also has the potential to build a quantitative evaluation platform that can be used in future chemical drug treatments.

SAFETY MARGIN IN ANUS-SAVING RESECTION FOR LOW RECTAL CANCER

The length and method of measurement of the safety-margin below the rectal cancer, being of the utmost importance for its prognosis, is still under debate.The following study was designed and done for its solution.Light microscopic examination was done on 83 resected rectal cancer specimens to assess the extent of intramural invasion towards the anus.By use of a ruler,the distance between the lower tumour margin and the resection line or the dentate line was measured when the specimen was:l. freshly resected,2.after fixing in 10% formalin, and 3.after being mounted in sections. The measurements were compared. By the same method,the distance between the lower tumor margin and the intended resectyion line was measured immediately before resection.It was compared with the measurement immediately after the resection.In 83 rectal cancer specimens, the extent of intramural infiltration toward the anus was:≤0.5 cm in 75 cases (90.4%).≥l cm in 2 cases which showed highly malignant carcinomas.These 2 cases, however,should not have been indicated for anus-saving resection.In 46 fresh specimens,the tumor-resection line distances gave an average of 2.7 cm.After fixing in 10% formalin, they became shortened by 0.7 cm. And, mouting in sections further shortened them by another 0.5 cm,giving a total shortening of l.2 cm.The tumor-resection line distance in 7 of the 11 fresh specimens resected by the Dixon's operation was shortened,though never more than o.5 cm immediately the operation.In performin ganus-saving resection for the low rectal cancer, after full isolation the rectum and stretching it slightly,≥3 cm of the rectum distal to the lower tumor margin should be resected.A safety margin of more than 2.5 cm is necessary in the fresh specimen.If formalin fixed specimen is measured, the safety margin should be ≥2 cm.

Gynaecological Cancers in HIV Positive and Negative Women—A Single-Center Retrospective Study (2008-2017)

Background/Objective: The association between Human Immunodeficiency Virus (HIV) and invasive cervical carcinoma is fully recognized. However, the effect of HIV and antiretroviral therapy on the morbidity and mortality of other gynaecological cancers have not been conclusively determined. Our study objective was to examine the effects of HIV on patient age at presentation, prevalence, and severity of the illness of various gynaecological cancers diagnosed in University of Nigeria Teaching Hospital (UNTH), Enugu over the period 2008-2017. Methods: This was a retrospective cross-sectional study of 224 patients who were managed in UNTH for different gynaecological malignancies. Ethical clearance was obtained from the Research Ethics Committee of the UNTH, Enugu. Data analysis was done with SPSS software with results expressed in descriptive statistics of simple frequency and percentage, and p-value set at Results: A total of 224 patients were studied. Twenty-five percent of HIV positive patients were aged 31 - 40 years at presentation compared to 12% of HIV negative patients. The commonest gynaecological cancer was cervical cancer with a higher proportion among the HIV-positive patients. While 32% of HIV negative patients presented at FIGO stages 1 - 2 Versus 8.3% of HIV positive patients, 58.3% and 33.3% of HIV positive patients presented at stages 3 and 4 respectively. Only 8.3% of HIV positive patients presented with ovarian cancer compared with 31% of HIV negative patients. A higher proportion of HIV positive patients presented with vulvar cancer (16.7%), but no endometrial or choriocarcinoma/GTD, compared with HIV negative patients. Conclusion: HIV positive clients present at an earlier age with more advanced disease, mostly cervical cancer of the squamous cell variety, with minimal non-AIDS defining cancers over the study period in UNTH, Enugu.

Effect of hypoxia-inducible factor-1α on proliferation and invasion of prostate cancer PC-3 cell in hypoxic situation

Objective We transfected recombinant expression plasmid of pcDNA3. 1-HIF-1α into prostate cancer cells, to research effect of HIF-1α on proliferation of prostate cancer cell PC-3. Methods We selected a stable expression cell line with G418 we selected by transfection

Can intravesical bacillus Calmette-Guerin reduce recurrence in patients with non-muscle invasive bladder cancer？ An update and cumulative meta-analysis

Approximately 70% of newly diagnosed bladder tumors are non-muscle invasive bladder cancer （NMIBC）. NMIBC accounts for approximately 80% of total bladder cancer cases. Bacillus Calmette-Guerin （BCG） instillation and maintenance is considered as the standard adjuvant treatment for superficial bladder cancer. A number of randomized studies have focused on the benefit of maintenance therapy following initial BCG induction. To provide further insights into the effect of intravesical instillation on recurrence in patients with NMIBC, we analyzed this relationship by conducting an updated detailed meta-analysis. Evidence suggested that adjuvant intravesical BCG with maintenance treatment is significantly effective for the prophylaxis of tumor recurrence in patients with NMIBC.

Inhibitory activities of plumbagin on cell migration and invasion and inducing activity on cholangiocarcinoma cell apoptosis

Objective: To investigate the cytotoxic, apoptotic and inhibitory activities on cell migration and invasion of plumbagin in the human cholangiocarcinoma(CCA) cell line(CL-6) in comparison with human embryonic fibroblast cell line(OUMS). Methods: Cytotoxicity activity was evaluated using MTT assay. Inhibitory effect on cell migration and invasion were investigated using label-free real-time cell analysis and QCM ECMatrix cell invasion chamber, respectively. Apoptotic activity was evaluated using flow cytometry and Cell Event? Caspase 3/7 assay. Results: Based on results of the cytotoxicity test in CL-6 cells, 50% inhibitory concentration(IC_(50), Mean±SD) values of plumbagin and the standard drug 5-fluorouracil were(24.00±3.33) and(1 036.00±137.77) μmol/L, respectively. The corresponding values for OUMS cells were(57.00±5.23) and(2 147.00±209.98) μmol/L, respectively. The selectivity index was 2.28. The inhibitory activities of plumbagin on cell migration and invasion were potent and concentration-dependent with IC_(50) of 25.0 μmol/L and complete inhibition at 25.0 μmol/L. Flow cytometry analysis showed that plumbagin at 12.5 μmol/L(half IC_(50)) induced CL-6 cell apoptosis(43.24% of control) through stimulation of caspase 3/7 activities. Complete cell apoptosis was observed at 12.5 μmol/L. Conclusions: The cytotoxic activity and inhibition of migration and invasion including apoptosis induction in the human CCA cell line(CL-6) suggest that plumbagin could be a promising candidate for CCA chemotherapeutics. However, its relatively low selective cytotoxic effect on CCA cells is a major concern.

Effects of Exogenous VEGF_(165)b on Invasion and Migration of Human Lung Adenocarcinoma A549 Cells

Vascular endothelial growth factor 165 （VEGF165）-mediated autocrine stimulation of tumor cells enhances the progression to a malignant phenotype. VEGF165b competes with VEGF165 and binds to vascular endothelial growth factor receptor （VEGFR）, resulting in inhibition of downstream signal transduction pathways. This study was designed to investigate the role of VEGF165b in the migration and invasion of human lung adenocarcinoma A549 cells. The full-length of VEGF165b was constructed and cloned into an expression plasmid （pVEGF165b）, and then transfected into A549 cells. Dimethylthiazolyl-2, 5-diphenyltetrazolium bromide （MTT） assay was used to detect the effect of VEGF165b on proliferation of transfected cells. Reverse transcription polymerase chain reaction （RT-PCR） was employed to examine the effect of VEGF165b on the expression of VEGF165 in transfected cells. Wound-healing assays were used to investigate the effect of VEGF165b on migration of transfected cells. Matrix metalloproteinase （MMPs） activity assay and in vitro invasion assay were used to determine the role of VEGF165b in invasion of transfected cells. There was no significant change in proliferation of A549 cells after transfection of pVEGF165b, but the expression of VEGF165, migration and invasion in A549 cells were inhibited. Furthermore, exogenous VEGF165b inhibited the activity of MMP9 in the supernatant of A549 cells and the subsequent invasion capacity of those cells. We therefore conclude that exogenous VEGF165b can inhibit the expression of VEGF165, as well as the migration and invasion of A549 cells, but has no effect on the proliferation of A549 cells.

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.