The Effect Of Regulator Of G Protein Signaling Proteins2,5on The Invasion And Migration Of The Ovarian Cancer Cell Lines Heya8

objectiveThe regulator of G-protein signaling5(RGS5) belongs to the family of GTPase activatorsthat terminate signal cascades initiated by extracellular mediators and G-protein-coupledreceptors.The RGS proteins have a variety of biological functions.The studies have reported that Gprotein signaling regulatory protein2,regulator of G protein signaling proteins5involved in thedevelopment, migration and invasion of malignant tumors. Associated with malignant tumors,in thisstudy we discuss the effect of RGS2, RGS5on the migration and invasion of epithelial ovariancancer.MethodsThis experiment was presented at the American University of Gerogia, Pharmacy colleageDr.Mandi Murph.The epithelial ovarian cancer cell lines expressing RGS2, RGS5respectively wereHeyA8-MDR pTet RGS2, HeyA8-MDR pTet RGS5. And not expressing RGS2, RGS5epithelialovarian cancer cell line HeyA8-MDR pTet Vector, the three groups of cells HeyA8-MDR pTetRGS2, HeyA8-MDR pTet RGS5, and the blank control group HeyA8-MDR Vector were screened ina medium with Taxol, G418.And then we put doxycycline in medium and culture the cells. TheCells grow along the sidewall, placed in37℃,5%C02incubator to culture; Cultured cells wereobserved,cell morphology and adherent conditions, until a cell density of80%-90%, with0.02%EDTA, trypsin solution after the cells were digested by centrifugation.We removed the culturemedium and adjusted the cell density to5×105/ml.The transwell chamber was put in24-well plates.The three groups of cell lines each containing5×105cells were inoculated in culture medium24well plate.Transwell (polycarbonate film thickness0.4um) chamber in the upper chamber.We added500ul culture medium in the upper chamber,and750ul in the lower chamber. We absorbed mediumin the two chambers after12hours. We added500ul cell culture solution Into the upper chamber, andwe added750ul cell culture solution and10%calf serum in to the lower chamber.Since the nutritionin the lower chamber was higher than the upper chamber, tumor cells in the upper chamber migratedto the lower chamber in order to obtain nutrition. We cultured cells routinly six to eight hours, then removed the transwell chambers and discarded the well of nutrient solution. We dyed the cells withcrystal violet, and washed them twice with PBS.We dried the small room at room temperature for10minutes, and compared with the control group by detecting the number of cells through into thelower chamber of the Transwell.reflecting the effect of RGS2, RGS5on the migration and invasionof epithelial ovarian cancer.Results(1) Compared with the control group, HeyA8-MDR pTet cells expressing RGS2gene havereduced invasive ability (t=0.0905, P <0,01, a statistically significant difference);(2) Compared with the control group, HeyA8-MDR pTet cells expressing RGS2gene havereduced metastatic ability (t=0.0086, P <0,01), a statistically significant difference);(3) Compared with the control group, HeyA8-MDR pTet cells expressing RGS5gene havereduced invasive ability (P <0,01, a statistically significant difference);(4) Compared with the control group, HeyA8-MDR pTet cells expressing RGS5gene havereduced metastatic ability (P <0,01, a statistically significant difference).Conclusion.Epithelial ovarian cancer cell lines,HeyA8-MDR pTet RGS2,expressing RGS2gene,theirmigration and invasion capacity reduced, HeyA8-MDR pTet with RGS5expressing RGS2gene,HeyA8-MDR pTet RGS5had reduced abilities of migration and invasion RGS5, indicating thatRGS2、RGS5can inhibit epithelial ovarian cancer cell migration and invasion.with the deepening ofthe study, the researchers could explore drugs targeting the RGS2, RGS5,providing new ideas andmethods for the treatment of epithelial ovarian cancer.