Sh-PLCε Improve the Efficacy of IL-2 in the Treatment of Renal Cell Carcinoma
YANG Jinxiao1, DUAN Limei1, FAN Jiaxin1, LI Ting1, FAN Yanru1, YUAN Hongling1, WU Xiaohou2, LUO Chunli1*
The purpose of this study was to investigate whether PLCε (phospholipase C ε) plays a role in the treatment of renal cell carcinoma with IL-2 (interleukin 2). We first built sh-PLCε cell line by transfecting LVsh-PLCε into renal caner cell 786-o. Cells were treated with the optimum concentration of IL-2 detected by MTT. Flow cytometry was used to detect apoptotic cells. DAPI staining was used to observe apoptotic bodies in cells. q-PCR and Western blot were used to detect Fas/FasL and Fas downstream molecules at mRNA level and protein level. Tumor cells and lymphocytes were co-cultured, then apoptotic cells were detected by flow cytometry. The results showed that the expression of Fas/FasL was decreased after knocking down PLCε, but the effect was reversed after IL-2 treatment. Detecting the downstream pathway of Fas, we found that FADD/cFlip/Traf2 signaling pathway was initiated in NC (negative control) group treated with IL-2 and the Fas downstream signaling pathway FADD/Caspase8/Caspase3 was initiated in sh-PLCε group after IL-2 treatment. The results of flow cytometry showed that the apoptotic rate was the highest in the group treated with IL-2 and sh-PLCε. This study demonstrated that sh-PLCε could improve the therapeutic effect of IL-2 on renal cell carcinoma by activating the downstream apoptotic signaling pathway of Fas and inhibiting the apoptotic inhibition signaling pathway.
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