Ultrasound-targeted microbubble destruction combined with dual targeting of HSP72 and HSC70 inhibits HSP90 function and induces extensive tumor-specific apoptosis
Ultrasound-targeted microbubble destruction combined with dual targeting of HSP72 and HSC70 inhibits HSP90 function and induces extensive tumor-specific apoptosis
复制标题
超声靶向微泡破坏结合 HSP72 和 HSC70 双重靶向抑制 HSP90 功能并诱导广泛的肿瘤特异性细胞凋亡
DOI：
10.3892/ijo.2014.2388
复制
发表时间：
2014-07-01
影响因子：
5.2
通讯作者：
Du, Lianfang
中科院分区：
文献类型：
--
作者：
Wang, Hanghui;Song, Yixin;Du, Lianfang
在体内将小干扰RNA（siRNA）特异性和有效地递送到癌细胞中仍然是一个主要障碍。在这项研究中，我们研究了超声靶向微泡破坏（UTMD）结合HSP 72和HSC 70在前列腺癌细胞系中的双重靶向是否提高了siRNA的特异性和有效的细胞摄取，抑制HSP 90功能并诱导广泛的肿瘤特异性凋亡。用siRNA寡核苷酸转染VCaP细胞。细胞活力测定用于评估UTMD的安全性。采用定量PCR和Western blotting方法检测HSP 70、HSP 90、caspase-8、caspase-3、PARP-1和裂解型caspase-3的表达。流式细胞仪检测细胞凋亡和转染效率。我们发现HSP 72、HSC 70和HSP 90在正常前列腺上皮细胞（RWPE-1）中不表达或表达较弱，而在前列腺癌细胞（VCaP）中变得均匀且强表达。VCaP和RWPE-1细胞表达非常低水平的caspase-8、caspase-3、PARP-1和裂解的caspase-3。UTMD联合H5 P72和HSC 70双靶向siRNA可提高siRNA的转染效率和细胞摄取，下调VCaP细胞HSP 70和HSP 90的mRNA和蛋白表达，上调主要凋亡标志物（PARP-1、caspase-8、caspase-3和cleaved caspase-3），诱导广泛的肿瘤特异性凋亡。细胞计数试剂盒-8测定显示HSP 72/HSC 70-siRNA沉默组中细胞活力降低。这些结果表明，UTMD与HSP 72和HSC 70的双重靶向的组合可以提高siRNA的特异性和有效的细胞摄取，抑制HSP 90功能并诱导广泛的肿瘤特异性凋亡，这表明一种新的、潜在的针对前列腺癌细胞的治疗策略的手段。
The specific and efficient delivery of small interfering RNA (siRNA) into cancer cells in vivo remains a major obstacle. In this study, we investigated whether ultrasound-targeted microbubble destruction (UTMD) combined with dual targeting of HSP72 and HSC70 in prostate cancer cell lines improve the specific and efficient cell uptake of siRNA, inhibit HSP90 function and induce extensive tumor-specific apoptosis. VCaP cells were transfected with siRNA oligo-nucleotides. Cell viability assays were used to evaluate the safety of UTMD. The expression of HSP70, HSP90, caspase-8, caspase-3, PARP-1 and cleaved caspase-3 were determined by quantitative PCR and western blotting. Apoptosis and transfection efficiency were detected by flow cytometry. We found that HSP72, HSC70 and HSP90 expression was absent or weak in normal prostate epithelial cells (RWPE-1), and became uniformly and strongly expressed in prostate cancer cells (VCaP). VCaP and RWPE-1 cells expressed very low levels of caspase-8, caspase-3, PARP-1 and cleaved caspase-3. UTMD combined with dual targeting of H5P72 and HSC70 siRNA impoved the efficiency of transfection, cell uptake of siRNA, downregulated HSP70 and HSP90 expression in VCaP cells on the mRNA and protein levels, and upregulated major apoptotic markers (PARP-1, caspase-8, caspase-3 and cleaved caspase-3), thus, inducing extensive tumor-specific apoptosis. The Cell Counting Kit-8 assay showed decreased cellular viability in the HSP72/HSC70-siRNA silenced group. These results suggest that the combination of UTMD with dual targeting of HSP72 and HSC70 may improve the specific and efficient cell uptake of siRNA, inhibit HSP90 function and induce extensive tumor-specific apoptosis, indicating a novel, potential means for targeting therapeutic strategy to prostate cancer cells.