Paclitaxel and B7-H6 knockdown inhibit HepG2 hepatocellular carcinoma cell viability and migration, while enhancing apoptosis and cell cycle arrest.
Hepatocellular carcinoma (HCC), the most frequent type of liver cancer, is a major source of cancer-related morbidity and mortality worldwide. B7-H6 is overexpressed in a variety of cancers including HCC and plays a key role in tumor biology and augments hepatoma cell proliferation, invasion, migration and cell-cycle progression. So, in this investigation we examined the role of B7-H6 silencing by specific siRNA in HepG2 cell line together with Paclitaxel treatment to reveal the potency of combining efficient targeted therapy with chemotherapy against tumor.
HepG2 cells were transfected with B7-H6-siRNA by the electroporation method, then treated with an appropriate dose of Paclitaxel determined by MTT. The consequence of B7-H6-siRNA transfection, Paclitaxel treatment and their simultaneous usage on the apoptosis, cell cycle and migration of HepG2 cells was evaluated by flow cytometry and wound-healing assay. Furthermore, the expression levels of Bax, Bcl-2, MMP9, B-Catenin, C-Myc and Cyclin D1 were also examined by quantitative real-time PCR.
Paclitaxel treated cells and B7-H6 suppressed cells and cells that received both of them became more sensitive to apoptosis and cell cycle arrest at the G2 phase and they showed reduced ability to migrate. Also, this therapeutic strategy had potency in reduction of BCL-2 as an anti-apoptotic gene and exaggeration of the apoptosis-involved gene Bax expression and reduction of other tumor associated genes' expression including MMP-9, B-Catenin, C-Myc and Cyclin D1.
These findings further support that Paclitaxel therapy in conjunction with B7-H6 suppression may present a potential option for successful cancer treatment.
HepG2 cells were transfected with B7-H6-siRNA by the electroporation method, then treated with an appropriate dose of Paclitaxel determined by MTT. The consequence of B7-H6-siRNA transfection, Paclitaxel treatment and their simultaneous usage on the apoptosis, cell cycle and migration of HepG2 cells was evaluated by flow cytometry and wound-healing assay. Furthermore, the expression levels of Bax, Bcl-2, MMP9, B-Catenin, C-Myc and Cyclin D1 were also examined by quantitative real-time PCR.
Paclitaxel treated cells and B7-H6 suppressed cells and cells that received both of them became more sensitive to apoptosis and cell cycle arrest at the G2 phase and they showed reduced ability to migrate. Also, this therapeutic strategy had potency in reduction of BCL-2 as an anti-apoptotic gene and exaggeration of the apoptosis-involved gene Bax expression and reduction of other tumor associated genes' expression including MMP-9, B-Catenin, C-Myc and Cyclin D1.
These findings further support that Paclitaxel therapy in conjunction with B7-H6 suppression may present a potential option for successful cancer treatment.
Authors
Alipour Alipour, Safdari Safdari, Khosravi Khosravi, Baghbani Baghbani, Karamali Karamali, Mehdizadehfar Mehdizadehfar, Baradaran Baradaran
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