Knockdown of TRPC5 in Regulating Ovarian Cancer Cells Resistance
Fu Qianyun1, Zhu Ruiyu2, Fang Luo1, Wu Miaolian1, Cai Zhibo1, Jin Jian2*
To investigate whether transient receptor potential 5 (TRPC5) is involved in affecting ovarian cancer cell resistance to paclitaxel (PTX), TPRC5-siRNA was transfected into A2780/PTX cells. MTT assay was used to detect the sensitivity of A2780/WT, A2780/PTX and A2780/PTX+siTRPC5 cells to paclitaxel. Real-time PCR (RT-PCR) and Western blot were used to detect the mRNA and protein expression levels of TRPC5 and Pglycoprotein (P-gp); cellular immunofluorescence to detect the expression levels of β-catenin, c-myc and Cyclin D1 proteins. The results of MTT, RT-PCR and Western blot showed that A2780/PTX cells (IC50=84.4 μmol/L)) were less sensitive to PTX than A2780/WT cells (IC50=1.98 μmol/L). Moreover, the mRNA and protein expression levels of TRPC5 and P-gp in A2780/PTX cells were significantly higher than those in wild cells. Furthermore, decreasing TRPC5 expression by using siTRPC5 in A2780/PTX cells significantly reduced P-gp expression and decreased sensitivity to PTX. In addition, immunofluorescence showed that TRPC5 knockdown in A2780/PTX cells reduced the nuclear translocation of β-catenin, and the expression of Cyclin D1 and c-myc was also significantly decreased. TRPC5 can enhance P-gp expression and drug resistance through Wnt/β-catenin signaling pathway, which suggest TRPC5 is involved in affecting the paclitaxel resistance in ovarian cancer cells.
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