The Mechanism Of CTRP6 Promoting Metastasis Of Ovarian Cancer

Objective: On the basis of RNAseq results of omental metastasis animal model of ovarian cancer,C1 q tumor necrosis factor related protein6,which was strongly expressed in the formation of omental metastasis microenvironment of ovarian cancer was screened out.The mechanism that CTRP6 promotes omental metastasis of ovarian cancer by regulating lipid metabolism through omental fat cells in vitro was further explored.Methods: The differentially expressed genes before and after the formation of omental metastasis of ovarian cancer were screened out.Using GO enrichment and analysis of the results of dynamic multi-point sequencing of omentum of ovarian cancer;the GEO data set (GES30587)and Kaplan Meier Plotter were used to analyze the expression of CTRP6 between primary and metastatic ovarian cancer and its relationship with the prognosis in ovarian cancer patients;stably overexpressed and knockout CTRP6 ovarian cancer cell lines were constructed by lentivirus,primary culture of preadipocytes,in co-culture system the proliferation of ovarian cancer cell was detect by growth curve,the adhesion of ovarian cancer cells to adipocytes was detect by the changes of ovarian cancer cells’ fluorescence signal,and Transwell was used to detect the migration of ovarian cancer cells;free fatty acid detection kit was used to detect the release of free fatty acids from adipocytes in co-culture system.And Western blot was used to verify the effect of CTRP6 on hormone sensitive lipase phosphorylation and AMPK pathway activation.Results:(1)By dynamic sequencing of d0,D1,D3,D10 and D21,it was found that D3 was the turning point before and after the formation of metastasis,and CTRP6 was strongly expressed in D3 and kept increasing;(2)GO enrichment analysis showed that the dynamic changes in the formation of omental metastasis of ovarian cancer mainly occurred in intercellular adhesion,immune regulation,inflammatory response and metabolism;(3)The analysis of GEO data set showed that the expression of CTRP6 in metastatic ovarian cancer was higher than that in primary ovarian cancer,and the high expression of CTRP6 was related to the poor prognosis in ovarian cancer patients(P < 0.05);(4)In vivo,the results showed that overexpression or knockdown of CTRP6 had no effect on the growth and migration of ovarian cancer cells.But in co-culture system,overexpression of CTRP6 could promote the growth,adhesion and migration of ovarian cancer cells(P < 0.05),and vice versa.The free fatty acid release and AMPK protein phosphorylation level of adipocytes were increased after co-culture with the overexpression of CTRP6 in ovarian cancer cells,and the adipose triglyceride lipase(ATGL)and hormone sensitive lipase(HSL)were activated.Conclusions:(1)According to the analysis,CTRP6 plays an important role in promoting omental metastasis of ovarian cancer in mouse model of intraperitoneal metastasis of ovarian cancer;(2)clinical data analysis showed that the expression of CTRP6 in metastatic ovarian cancer is higher than that in primary ovarian cancer and is related to prognosis in ovarian cancer patients;(3)By activating the phosphorylation of AMPK at th172 site,CTRP6 promoted the expression of ATGL and the phosphorylation level of HSL in adipocytes,promoted the release of free fatty acids in adipocytes,and thus participated in the growth,adhesion and migration of ovarian cancer cells.