Silencing ANO9 increases the radiosensitivity of pancreatic cancer cell AsPC-1 / 中华放射肿瘤学杂志

ABSTRACT

Objective To evaluate the effect of ANO9 on the radiosensitivity of pancreatic cancer cell AsPC-1,aiming to provide new targets for clinical radiotherapy of pancreatic cancer.Methods Western blot was performed to detect the expression of ANO9 in pancreatic cancer cell lines (BxPC-3,PANC-1,AsPC-1)and normal pancreatic cell line (HPNE).The AsPC-1 cell line with stable silencing ANO9 was constructed by using lentivirus and validated by Western blot.MTT assay was adopted to detect the cell viability of AsPC-1 with stable silencing ANO9 after irradiation.Colony formation assay was conducted to evaluate the effect of silencing ANO9 upon the radiosensitivity of AsPC-1 cells.Western blot was performed to assess the effect of ANO9 silencing on the expression of EGFR/ERK signaling protein.Results The expression levels of ANO9 were significantly up-regulated in three pancreatic cancer cell lines compared with that in the normal pancreatic cell line HPNE (t =7.426,5.543,11.850,all P＜0.05).After silencing ANO9,the expression level of ANO9 protein was significantly down-regulated than that in the control group (t =9.670,P＜0.05).The AsPC-1 cells with stable silencing ANO9 were successfully constructed.The sensitivity of AsPC-1 cells to irradiation was significantly increased after silencing ANO9,and the sensitivity enhancement ratio was 1.566.The expression levels of EGFR/ERK signaling proteins (EGFR and p-ERK 1/2) were significantly downregulated after silencing ANO9 (t =7.949,13.160,both P＜ 0.05).Conclusions Silencing ANO9 can significantly increase the sensitivity of AsPC-1 cells to radiotherapy,which is probably associated with the inhibition of EGFR/ERK signaling transduction.ANO9 might be a new therapeutic target for preventing the progression of pancreatic cancer.