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@#[Abstract] Objective: To construct and verify the anti-tumor activity of chimeric antigen receptor (CAR) modified NK-92 cells (CAR-NK-92 cells) targeting prostate stem cell antigen (PSCA) in cervical cancer. Methods: Lentiviral vector expressing CAR targeting PSCA was constructed, and PSCA CAR-NK-92 cells were obtained by lentivirus transfection. The expression of PSCA in human cervical cancer cells was determined by Flow cytometry and Western blotting. The killing effect of PSCA CAR-NK-92 cells against cervical cancer cells was verified by co-incubation of effector and target cells in vitro, and the tumor inhibitory ability of PSCA CAR-NK-92 cells was verified with the nude mice xenograft model in vivo. Results: PSCA CAR-NK-92 cells were successfully constructed. PSCA was highly expressed in human cervical cancer Hela and MS751 cells (all P<0.01). In vitro co-incubation results showed that PSCA CAR-NK-92 cells could lyse PSCA+ cervical cancer transplanted tumor in a dose-dependent manner. In vivo anti-tumor data showed that PSCA CAR-NK-92 cells significantly inhibited the growth of cervical cancer cells compared with NK-92 cells transfected with vehicle vectors (P<0.01). In addition, PSCA CAR-NK-92 cells could effectively infiltrate tumor tissues and promote the secretion of anti-tumor cytokines TNF-α and IFN-γ (all P<0.01). Conclusion: The CAR-NK-92 targeting PSCA shows good anti-tumor effect on PSCA+ tumor cells both in vitro and in vivo, and has potential to be a therapeutic strategy for cervical cancer.
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PURPOSE: Half of the world's gastric cancer cases and the highest gastric cancer mortality rates are observed in Eastern Asia. Although several genome-wide association studies (GWASs) have revealed susceptibility genes associated with gastric cancer, no GWASs have been conducted in the Korean population, which has the highest incidence of gastric cancer. MATERIALS AND METHODS: We performed genome scanning of 450 gastric cancer cases and 1,134 controls via Affymetrix Axiom Exome 319 arrays, followed by replication of 803 gastric cancer cases and 3,693 healthy controls. RESULTS: We showed that the rs2976394 in the prostate stem cell antigen (PSCA) gene is a gastriccancer-susceptibility gene in a Korean population, with genome-wide significance and an odds ratio (OR) of 0.70 (95% confidence interval [CI], 0.64 to 0.77). A strong linkage disequilibrium with rs2294008 was also found, indicating an association with susceptibility. Individuals with the CC genotype of the PSCA gene showed an approximately 2-fold lower risk of gastric cancer compared to those with the TT genotype (OR, 0.47; 95% CI, 0.39 to 0.57). The effect of the PSCA gene on gastric cancer was more prominent in the female population and for diffuse type gastric cancer. CONCLUSION: Our result confirmed that the PSCA gene may be the most important susceptibility gene for gastric cancer risk in a Korean population.
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Female , Humans , Exome , Asia, Eastern , Genetic Variation , Genome , Genome-Wide Association Study , Genotype , Incidence , Linkage Disequilibrium , Mortality , Odds Ratio , Prostate , Stem Cells , Stomach NeoplasmsABSTRACT
Objective: To evaluate the association between rs2294008 C>T polymorphism of prostate stem cell antigen (PSCA ) gene and the risk of gastric cancer in East Asian by using Metaanalysis. Methods: A computer-based online search was performed in PubMed, Web of Science, Cochrane Library, Google Scholar, China National Knowledge Infrastructure, Chinese BioMedical Literature database, VIP database and Wanfang database. The case-control studies about the association between PSCA rs2294008C>T polymorphism and gastric cancer in East Asian before June 6, 2016 were collected. The literatures were screened according to the inclusion and exclusion criteria, and the data were extracted after the quality analysis. The Meta-analysis was performed by STATA 14.1 software, and the odds ratio (OR ) and 95% confidence interval (CI ) were calculated. Furthermore, the subgroup analysis and Metaregression analysis were used to evaluate the confounding factors, and the sensitivity analysis and publication bias test were performed. Results: A total of 13 case-control studies contained 13 667 patients and 11 868 health controls were included. Meta-analysis showed that the carriers with PSCA rs2294008 CT and TT genotypes had significantly higher risk of gastric cancer as compared with other genotype (OR = 1.68, 95% CI: 1.43-1.96). The polymorphism distribution of PSCA rs2294008 was significantly different among Chinese, Japanese and Korean populations (P = 0.001). Subgroup analyses showed that the risk of gastric cancer on the carriers with PSCA rs2294008 CT and TT was significantly increased in Chinese (OR = 1.36, 95% CI: 1.23-1.49, P = 0.004), Japanese (OR = 2.61, 95% CI: 2.22-3.07, P = 0.001) and Korean populations (OR = 2.04, 95% CI: 1.28-3.27, P = 0.012) compared with rs2294008 CC. The sub-group analyses also showed that there was no significant difference in the risk of gastric cancer between the controls from hospitals (OR = 1.74, 95% CI: 1.23-2.44) and ones from communities (OR = 1.64, 95% CI: 1.40-1.93). The Meta-regression found the differences in countries and control sources were not the major contribution to the heterogeneity in this analysis (P = 0.163, P = 0.381). The Begg's funnel plots showed no publication bias (corrected P = 0.053), and the sensitivity analysis confirmed the data were steady (OR = 1.54, 95% CI: 1.45-1.64). Conclusion: The polymorphism of PSCA rs2294008 C>T is closely associated with the risk of gastric cancer in East Asia.
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Background:Gastric cancer is the result of comprehensive interactions among Helicobacter pylori(Hp)infection, environmental factors and host genetic factors. It has been demonstrated that rs2294008 polymorphism in prostate stem cell antigen(PSCA)gene is associated with increased risk of non-cardia gastric cancer. Aims:To investigate the relationship between PSCA rs2294008 polymorphism and precursors of gastric cancer. Methods:A total of 398 patients with gastric intestinal metaplasia( n = 328)and intraepithelial neoplasia( n = 70)from Nov. 2009 to Nov. 2015 at the Qingdao Municipal Hospital were enrolled,and 416 healthy subjects were served as controls. Genotype of PSCA rs2294008 was determined by direct DNA sequencing of PCR products,and Hp infection was examined by rapid urease test. Results:Significant difference in frequencies of CC,CT and TT genotypes of PSCA rs2294008 was observed between case and control groups(P = 0. 011);the frequency of TT genotype was significantly higher in case group than in control group (16. 3% vs. 9. 4% ,P = 0. 003). Compared with individuals carrying CC genotype,TT genotype carriers were found to be associated with a higher risk of precursors of gastric cancer(oR = 1. 840,95% CI:1. 174 ~ 2. 886). Taken individuals negative for Hp infection and carrying C allele(CC + CT)as reference,risk of precursors of gastric cancer was significantly increased by Hp infection(oR = 2. 389,95% CI:1. 799 ~ 3. 173)and Hp infection in combination with TT genotype (OR = 3. 335,95% CI:1. 935 ~ 5. 749),whereas TT genotype alone only slightly increased the risk(OR = 1. 783,95%CI:0. 900 ~ 3. 530). Conclusions:PSCA rs2294008 polymorphism is significantly associated with susceptibility to precursors of gastric cancer and Hp infection might further increase the risk in TT carriers.
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Methods: Twenty-five healthy male C57BL/6 mice were randomly divided into 5 groups (5 each): PSCA, HSP, PSCA+HSP, PSCA-HSP and control group. Mice in the first 4 groups were vaccinated with the corresponding proteins, and those in control group were faked with injection of phosphate buffer saline (PBS). After immunization with recombinant proteins, the PSCA-specific cellular immune responses were monitored with ELISPOT, intracellular cytokine staining assay, and flow cytometry, and ELISA assay was used to detect humoral immune responses. The tumor growth and survival of vaccined mice were observed.
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Recently,some researches have been focused on prostate stem cell antigen and related cancers.Researches have reveled that prostate stem cell antigen is overexpressed in prostate cancer,bladder cancer and some malignant tumors in digestive system,and is highly associated with tumour genesis and progress.This review is to show the recent researches on association between prostate stem cell antigen and bladder cancer.
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Objective To investigate the role of the expression of prostate stem cell antigen(PSCA)in the development of perineural invasion in pancreatic carcinoma.Methods The histopathologic and immunohis-tochemical(SABC)studies were performed on 80 patients with pancreatic carcinoma.The overall incidence of PSCA expression,perineural,lymphatic and vascular vessel invasion were counted to investigate the relationships among them.Results Perineurel invasion was positively correlated with lymphatic and vascular vessel invasion (P<0.01).A positive corelation Was found between tumor PSCA expressioa(53 cases)and the perineurel in-vasion(66 cases).A definite correlation Was also found between cancer differentiation and PSCA tyxplres-sion.Conclusion PSCA is one of the most important molecules and may play a role as a "navigating" and " docking" molecule in the developmeat of perineural invasion.
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Objective To establish a mouse model of prostate cancer expressing human PSCA for the development of new anti-tumor drugs or vaccines. Methods The total RNA of DU145 cells,a human prostate cancer cell line,was isolated by using TRIzol reagent according to the (RT-PCR),the first-strand cDNA was synthesized using the SuperScript First-Strand synthesis system. The human PSCA gene was amplified with the primers and cloned into the plasmid pcDNA3.1 to generate pcDNA-PSCA. DNA sequencing was used to confirm the constructs. The mouse prostate tumor cell line RM-1 cells,syngeneic to C57BL/6,were transfected with pcDNA-PSCA plasmids followed by selection using G418. RT-PCR analysis was performed to examine the validity of the constructs. Expression of PSCA on the cell surface was determined by staining with anti-PSCA antibody,and the anti-PSCA antibody was detected using an FITC-conjugated goat anti-rabbit IgG antibody,and analyzed by flow cytometry. 4-6-week-old male C57BL/6 mice purchased from the Laboratory Animals Center were inoculated with different amounts of RM-PSCA cells to search for suitable cell population which can form tumor in mouse,and the mice were monitored twice a week. The growth and the survival time of mice were measured,respectively. The tumor volume was measured by vernier caliper according to the formula:V=0.5a×b~2,where a and b are the long and short diameters of the tumor,respectively. Results The plasmid pcDNA-PSCA was successfully constructed and the PSCA was successfully expressed in RM-PSCA 7~# and RM-PSCA 28~# cells by RT-PCR and confirmed by flow cytometry. 1×10~5 RM-PSCA cells were sufficient to get tumor growth in 100% of inoculated mice. The tumor grew quickly and the volume of the tumor reached 12000 mm~3 within 34 days. All the mice died within 40 days and their mean survival time was 37 days. Conclusion A PSCA-expressing tumor model in mice has been successfully established. It can be used to evaluate the activities of drugs or vaccines.
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Objective To prepare monoclonal antibody against Prostate Stem Cell Antigen(PSCA) and explore the inhibitory effects of Anti-PSCA mAb in treatments of human prostate cancer xenografts in mice. Methods Balb/c mice were immunized by PC-3 cell line. After fusing and screening, the anti-PSCA McAb’s characterizations were determined. Solid tumors in mice were produced by subcutaneous injection with PC-3 cells in the flanks of the mice.We picked out 10 mice bearing human prostate cancer xenografts.They were divided into a treatment group (n_1=5) and a control group (n_2=5).200 ?g Anti-PSCA mAb was injected into abdominal cavity of each mouse of the treatment group and PBS for them of the control group.Anti-PSCA mAb and PBS were administered once every three days for consecutive three times.The mice survival conditions of two groups were recorded during 5 weeks.The serum PSA, the tumor weights and dimensions of survived mice were measured.The tumor volume inhibition rate was calculated. T-test was performed to compare differences of PSA in serum,tumor weights and volumes between the treatment and control groups.Routine pathological slides of tumor tissue were observed under light microscope to evaluate the range of tumor tissues damaged by Anti-PSCA mAb. Ultrastructure was observed with transmission electron microscope. Results A marine McAb was produced, which raised against PSCA,belonged to IgG1 subclass. The average PSA serum level of the two groups were(3.28?0.55)ng/ml and(7.26?0.43)ng/ml.The weights of tumors of the two groups were(0.95?0.17)g and(3.08?0.18)g.The volumes of them were(164.59?14.08)mm3 and(548.49?19.79)mm3.There are remarkable differences between the treatment group and the control group(P
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Objective To investigate the expression of prostate stem cell antigen(PSCA) in human prostate cancer (PCa) specimens and its possible correlations with pathological grade and clinical stage of PCa. Methods Immunohistochemical (IHC) and in situ hybridization (ISH) analyses of PSCA expression were simultaneously performed on paraffin-embedded sections from 20 cases of benign prostatic hyperplasia (BPH),20 cases of prostatic intraepithelial neoplasm (PIN) and 40 cases of (PCa) tissues.The levels of PSCA protein mRNA expression were semiquantitatively scored by assessing both the percentage and intensity of PSCA-positive staining cells in the specimens,and then were compared among PCa,BPH and PIN tissues. Results The moderate to strong positive rate of expression of PSCA protein and mRNA in PCa,BPH and PIN tissue was 85%,20% and 35%,respectively.Significant difference of expression was found between PCa and BPH,PIN samples (P0.05).The expression level of PSCA increased with higher Gleason score and worsening clinical stage (P