Effects Of Mesothelin On Transplanted Tumor Growth Of Human Epithelium Ovarian Cancer And Gene Therapy In Nude Mice

Objective:Stablely transfected cell lines SKOV3- MLSN- cDNA and SKOV3-MLSN-shRNA, the mesothelin gene of which was up-regulated and down-regulated, were developed by chronic lentivirs mediated RNAi. Ovarian cancer nude mice model was established using the cells, and the effect of mesothelin on body weight and abdominal circumference was observed. The effect of mesothelin in epithelium ovarian cancer and gene therapy in vivo was studied.Methods: 1 Animal model of nude mice are established Femal BALB/c nude mice, 4-6weeks old and 13.5-16.5g weight, live in clearning environment. SKOV3, SKOV3- MLSN-cDNA, SKOV3-MLSN-shRNA, SKOV3/neg cells were injected into the peritoneal cavity of nude mice. After 7-8 days,The body weight and abdominal circumference of nude mice obviously increased.2 Experiment groupTest one: SKOV3, SKOV3-MSLN-cDNA, SKOV3-MSLN-neg, SKOV3-MSLN-shRNA cells were injected into the peritoneal cavity of nude mice; Test two: tumor cells SKOV3 and LV-MSLN-cDNA,LV-MSLN-neg, LV-MSLN-shRNA, PBS were injected at the same time; Test three: SKOV3 and Lentivirus were injected at the frist day, Lentivirus were injected every other day; Test four: only lentivirus were injected. Weight, distribution and ascites of transplanted tumor were measured after 14 days.3 Western Blotting and Immunohistochemical method were used to measure expression of MSLN in test three.4 Investigation of changes in nude mice organs in test four by nake eye and HE chromoscopy.Results: 1 Transplanted tumor of SKOV3-MSLN-shRNA cells were suppressed obviously in nude mice compared with the control cells (P<0.05).2 In test two, the growth of SKOV3 were not suppressed obviously by LV-MSLN-shRNA.3 But in test three, suppression was obvious by LV-MSLN-shRNA, compared with the control cells.4 Western Blotting results: Interfere efficiency in test 3 treated by LV-MLSN-shRNA and LV-MLSN-neg chronic lentivirs is 75.6% and 8.9%, compared with control group treated by PBS. The expression of MSLN in treated by LV-MLSN-shRNA chronic lentivirs was significantly lower than the negative control group and the blank control group (P<0.05). The negation control group and the blank control group were no siginificant difference in the interfere efficiency (P>0.05).5 Expression of MSLN in tumor issue by Immunohisto -chemical method results: MSLN localizes on membrane of tumor cells. The HIS value is 3.20±0.33 in the group treated by LV-MSLN-shRNA, and is 7.42±0.53 treated by LV-MSLN-neg, and is 7.88±0.29 in control group treated by PBS. The expression of MSLN in treated by LV-MLSN-shRNA chronic lentivirs was significantly lower than the negative control group and the blank control group (P<0.05). The negation control group and the blank control group were no siginificant difference in the interfere efficiency (P>0.05).6 Three kinds of lentivirus have no toxic effec on nude mice.Conclusion: 1 The metastsis and implation of SKOV3 in abdominal cavity were effected by MSLN gene. If MSLN is low-expressed, the number and weight of xenografts will be decreased. Thus the metastsis and implation of ovarian cancer can be delayed.2 Lentivirus mediated RNA interference of mesothelin suppressed the transplanted tumor growth of human epithelium ovarian cancer. It will be a powerful strategy for cancer gene therapy.3 Toxicin was not found in experimental animals treated by Chronic lentivirs, so Chronic lentivirs therapy is a safe and reliable method.