Effect Of Overexpressing LIMK1 On Inhibiting Gastric Cancer In Nude Mice Xenografts By Rosiglitazone

Objectives :By constructing stable LIMK1 overexpression of MGC-803 cell lines to observe the effect of gastric cancer cells in nude mice xenografts with LIMK1 and rosiglitazone(Rosiglitazone, ROS).Methods :Constructing the LIMK1 overexpression viral vector systems which by lentivirus-mediated to infected MGC-803 cells and screening stably overexpressing LIMK1 cells. To identify the stable cell lines By q PCR and Western blot. Gathering OE group, NC group and control group,the three groups of cells were inoculated into nude mice forelimb armpit to observe tumorigenesis situation on nude mice. When the xenograft growing into100mm3 beginning to use medicine, Firstly, They were divided into two groups in the three groups each randomly, a total of six groups, each group contains of eight xenograft nude mice. Experimental groups were given rosiglitazone 100 mg / kg / 2d,and the control groups received normal saline.Measured the tumor volumes and weighed the mice every 6 days,then draw ing the growth curve of xenograft tumor. Observated morphological changes of the xenograft tumor by HE stain, The expression of LIMK1 protein were detected by immunohistochemistry and Western blot after being treated by ROS in nude mice, Finally, statistical analysis was performed.Results: 1. successfully constructed stably overexpressing LIMK1 of MGC-803 cell line.With lentivirus-mediated LIMK1 overexpression viral vectors to infect MGC-803 cells, Green fluorescent cells were detected prompting that MGC803 cells had been infected the virus successfully. The cells can be observed stably expressing green fluorescence, After being screened by puromycin. The q PCR and Western blot experiments showed that the LIMK1 overexpression group of m RNA and protein expression level higher than empty vector group(P<0.05),However,The Transfected with empty vector group MGC803 cells compared with the control group, the protein expression levels of LIMK1 have no statistically significant difference(P> 0.05).2. Effect of ROS and overexpression of LIMK1 on MGC803 cells in nude mice xenografts experiments.(1) Effect on tumor growth: The IMK1 overexpression group of size and weight of the control group significantly higher than NC group,and the ROS drenching group of size and weight significantly lower than drenching group.The perfusion group had some inhibitory rate afte being Calculated.(2) Effect on tumor morphology: No perfusion group has a larger cell atypia, more interstitial cells, deeper nuclear staining, multi-nuclearcytoplasmic ratio, cell disorder, comparing with the ROS drenching group, of which LIMK1 overexpression group is the most obvious.(3) Effect of tumor protein expression: Each group has LIMK1 protein expression in transplanted tumor, In 40 X objective observation by contrast, In group C, the expression was(LIMK1 overexpression) the most obvious, No perfusion group(A, B, C) of LIMK1 expresss more protein compared with ROS perfusion group(D?E?F).Conclusion:1. Overexpression of LIMK1 may promote the growth of Xenografts in nude mice.2. Rosiglitazone may inhibit the transplanted tumor growth of Xenografts in nude mice.3.LIMK1 may be a potential target of the inhibition effects of rosiglitazone on the proliferation, migration and invasion in MGC803 cells.