MicroRNA-145 Inhibited The Proliferation Of Ovarian Cancer Cells In Animal Experiments

Objective:We set an experiment of tumor-transplantation through building nude mice model which can express shRNA of microRNA-145 stably in ovarian cancer cell line(SKOV3-miR145-CMV-RFP).We wanted to investigate how microRNA-145 influence the proliferation of ovarian cancer cell in vivo.Methods:We built miR-145-CMV-RFP-SKOV3’s stable cell line and designed primer sequences of miR-145shRNA. Then we synthetized sequence,connected it with carriers,packed slow virus, transfected SKOV3 cells using slow virus and chose the stable cell line which was transfected by SKOV3 cells。We examined the SKOV3 cells to validate the degree of integration for stably expressing miR-145.Then we built nude mouse transplantation tumor model. The mice were divided into the experimental group,the negative control group and blank control group at random. The experimental group was transfected by SK-OV-3-rfp-shRNA,the negative group was transfected by SK-OV-3-rfp and the blank group was transfected by SK-OV-3. We planted 0.1ml tumor cells suspension on back subcutaneous of each nude mouse and observed the tumors.We measured the length and width every three days,and calculated the volume of the tumors,then drawn a tumor curve. After 30 days,we weighed the mice,then killed the mice by taking off necks.We stirpped the tumors completely and took photos to record the size of the tumors.The tumors were detected by Western Blot for expression of protein MUC1.Results:1. The miR-145-CMV-RFP-SKOV3’s stable cell line was successfully built.Its degree of integration is high:The integrated degree of SK.OV3 cells which were transfected by LvX-MOCK-CMV-RFP has reached more than 90%, and the degree of SKOV3 cells transfected by LvX-miR-145-CMV-RFP was also more than90%.It made SKOV3 cells to express miR-145 stably。2. The inhibiting effect of miR-145-CMV-RFP-SKOV3 in tumor-transplantion in nude mice which were transfected by SKOV3:We observed the tumors’growth and measured the volume of the tumors every three days and drawn the tumor curve.Compared with the blank group which were transfected by SK-OV-3, the experiment group which were inoculated by SK-OV-3-rfp-shRNA were growing slowly and the volume of the tumor was significant reduced (P<0.01).Compared with the blank group,the negative group which were transfected by SK-OV-3-rfp has shown reduced volume of the tumor,but there is no statistical significance (P>0.05).The outcomes of Western Blot demonstrate that:Compare with the blank group, the expression of protein MUCl in experiment group’s tumor tissues were falled clearly (P<0.01). Compare with the blank group, the expression of protein MUCl in negative group’s tumor tissues had no obvious differences (P>0.05).It turned out that miR-145may regulate the expression of MUCl to inhibite the proliferation of ovarian cancer SK-OV-3 cells.Conclusions:miR-145-CMV-RFP-SKOV3 has obvious inhibiting effect on the proliferation of SKOV3 cells in nude mice which may be related to the regulation of protein MUCl.