Roles Of Cancer Associated Fibroblasts In The Malignant Behavior Of Human Ovarian Cancer

Objective: To extract Cancer-associated fibroblasts (CAFs) fromhuman ovarian cancer tissues of patients with different kinds of histologicaltypes for studying the roles of CAFs in the malignant behavior of ovariancancer.Methods:1. CAFs were isolated from tumor lesions in human ovariancancer specimens. Immunocytochemistry was used to identify CAFs andNFs by quantifying the expression of α-SMA.2.The CAFs density inprimary and metastatic tumors were measured throughimmunohistochemistry assay. Moreover, the quantification of SDF-1secreted from CAFs in primary and metastatic tumor sites was also testedby ELISA.3. Transwell invasion assay was used to investigate the differentinvasiveness of CAFs from tumor cells, both of which were isolated fromthe same ovarian cancer specimen.4. Cell migration of SKOV3,OVCAR-3and HEY were observed in co-cultured with CAFs by transwell migrationassay.5. After treated with CDDP, the apoptosis of CAFs and tumor cellswere evaluated by single cell gel assay. And immunofluorescence assay was used to test the expression of α-SMA in CAFs with/without CDDPtreatment.Results:1.CAFs were identified by the high level α-SMA, which wasmuch less expressed in NFs.2. CAFs density was found to be rich inmetastatic lesions compared with in primary tumor site of human ovariancancer. However, there was no significant difference of SDF-1productionfrom CAFs between primary and metastatic lesions.3. More CAFs wereobserved to invade through matrigel than tumor cells by transwell invasionassay.4.When co-cultured with CAFs, SKOV3and HEY cells showed asignificant increase in cell migration ability compared with those culturedalone（P<0.01）.5.As expected, obvious apoptosis was observed in tumorcells other than in CAFs when treated with CDDP（P<0.01）. Furthermore,higher α-SMA expression was detected in CAFs treated with CDDP thanthose without（P<0.01）.Conclusions: CAFs might be involved in the malignant metastasis ofhuman ovarian cancer through promoting cell migration of tumor cells.Moreover, the resistance to cytotoxic agents might make the tumormicroenvironment more conductive to tumor cells, even though theunderlying mechanism needs further identifying.