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目的探讨蛋白激酶D3(protein kinaseD3,PKD3)是否会影响依托泊甙对激素抵抗性前列腺癌(hormonal refractory prostate cancer,HRPC)的生长抑制。方法在HRPC细胞系DU-145中瞬时转染PKD3的siRNA,Western blot检测细胞中内源性PKD3的表达变化;siRNA转染48h后,分别以0、30、70、100、300μmol/L的依托泊甙处理对照siRNA、siRNA-PKD3-1、siRNA-PKD3-2转染的DU-145细胞24h,并以细胞活力测定观察敲定内源性PKD3的表达有无影响依托泊甙对DU-145的生长抑制;利用PKC单一抑制剂、PKC-PKD双重抑制剂处理DU-145细胞,以观察二者是否增强DU-145对依托泊甙的敏感性。结果 Westernblot证实siRNA-PKD3的转染敲低DU-145细胞内源性PKD3的表达;细胞活力测定表明,在非特异性siRNA(si-CTL)转染的DU-145细胞中,依托泊甙剂量依赖地抑制DU-145细胞的生长,而敲低PKD3的表达不仅可显著抑制DU-145的生长(P<0.01),还可显著增强依托泊甙对DU-145生长的抑制(P<0.01);Go6976(PKC-PKD的双重抑制剂)可明显抑制前列腺癌DU-145细胞生长(P<0.01),而Go6983(单一的PKC抑制剂)则仅对DU-145的生长起微弱抑制作用。结论 PKD3的表达和活性可增强前列腺癌DU-145细胞对依托泊甙的敏感性,提示可作为抗前列腺癌药物设计的潜在靶点之一。
Objective To investigate whether protein kinase D3 (PKD3) may affect the growth inhibition of etoposide on hormone-refractory prostate cancer (HRPC). Methods PKD3 siRNA was transiently transfected into HRPC cell line DU-145 and the expression of endogenous PKD3 was detected by Western blot. After transfected with siRNA for 48h, the cells were treated with 0, 30, 70, 100, 300μmol / L The effect of etoposide on the expression of DU-145 in DU-145 cells transfected with control siRNA, siRNA-PKD3-1, and siRNA-PKD3-2 was observed by cell viability assay for endogenous PKD3 expression Growth inhibition. DU-145 cells were treated with PKC single inhibitor and PKC-PKD double inhibitor to observe whether DU-145 enhanced the sensitivity to etoposide. Results Western blot confirmed that the transfection of siRNA-PKD3 knocked down the expression of endogenous PKD3 in DU-145 cells. The cell viability assay showed that the dose dependence of etoposide in DU-145 cells transfected with nonspecific siRNA (si-CTL) Inhibited the growth of DU-145 cells. However, knockdown of PKD3 expression not only inhibited the growth of DU-145 significantly (P <0.01), but also inhibited the growth of DU-145 significantly (P <0.01). Go6976, a dual inhibitor of PKC-PKD, significantly inhibited the growth of prostate cancer DU-145 cells (P <0.01), while Go6983 (a single PKC inhibitor) only slightly inhibited the growth of DU-145 cells. Conclusions The expression and activity of PKD3 can enhance the sensitivity of etoposide to prostate cancer DU-145 cells, suggesting that PKD3 may be one of the potential targets for anti-prostate cancer drug design.