Huang Wan-lan, Li Xu-yuan, Wang Hong-biao, Lin Wen, Lin Wen-zhao, Lin Sui-ling, Lin Ying-cheng
Department of Medical Oncology, Cancer Hospital, Shantou University Medical College, Shantou 515031, China.
Zhonghua Yi Xue Za Zhi. 2012 Apr 24;92(16):1133-7.
To explore the in vitro effects of anti-proliferation and apoptosis-inducing with different sequence regimens of zoledronic acid plus paclitaxel in human nasopharyngeal carcinoma cell line HNE1 so as to explore the optimal sequence regimen of these two drugs and related mechanism.
The cytotoxic effects of different sequence schemes of zoledronic acid plus paclitaxel on HNE1 cells were detected by methyl-thiazol-tetrazolium (MTT) assay. Annexin V-FITC/PI double staining flow cytometry (FCM) and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay were used to measure the effects of zoledronic acid plus paclitaxel upon apoptosis. The expressions of mRNA of Bcl-2, Bax, Caspase3 and Caspase9 gene were detected by real-time quantitative-polymerase chain reaction (PCR) and protein was detected by Western blot.
All experiment groups enhanced the effect of anti-proliferation by MTT assay (P < 0.05); the treatment of zoledronic acid followed by paclitaxel was superior to the other two regimens (P < 0.05). As detected by FCM, the early apoptotic rate of control group was 2.59% ± 0.28% and the experiment groups were 13.89% ± 0.69%, 11.73% ± 0.54%, 23.97% ± 0.68%, 10.45% ± 0.16% and 8.59% ± 0.74% respectively (P < 0.05). TUNEL assay detected the late apoptosis of HNE1 cells and the experiment groups enhanced the effect of apoptosis-inducing (P < 0.05). The treatment of zoledronic acid followed by paclitaxel was superior to the other regimens (P < 0.05). Such an effect was due to the down-regulation of anti-apoptotic protein Bcl-2 and up-regulations of pro-apoptotic proteins Bax, Caspase3 and Caspase9 at the expression levels of mRNA and protein. There was a greater regulation in the group of zoledronic acid followed by paclitaxel.
Zoledronic acid can enhance the in vitro effects of anti-proliferation and apoptosis-inducing for paclitaxel on HNE1 cell. The treatment of zoledronic acid followed by paclitaxel may be the optimal regimen. Synergistic induction of apoptosis is via the effects of Bcl-2 family and through the mitochondrial pathway.
探讨唑来膦酸联合紫杉醇不同给药顺序方案对人鼻咽癌HNE1细胞株的体外抗增殖及诱导凋亡作用，以探寻两种药物的最佳给药顺序方案及相关机制。
采用甲基噻唑基四氮唑蓝（MTT）法检测唑来膦酸联合紫杉醇不同给药顺序方案对HNE1细胞的细胞毒性作用。采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶（Annexin V-FITC/PI）双染流式细胞术（FCM）和末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记（TUNEL）法检测唑来膦酸联合紫杉醇对细胞凋亡的影响。采用实时定量聚合酶链反应（PCR）检测Bcl-2、Bax、Caspase3和Caspase9基因mRNA的表达，采用蛋白质免疫印迹法检测蛋白质表达。
MTT法检测结果显示，各实验组均增强了抗增殖作用（P<0.05）；唑来膦酸后用紫杉醇的治疗方案优于其他两种方案（P<0.05）。FCM检测结果显示，对照组早期凋亡率为2.59%±0.28%，各实验组分别为13.89%±0.69%、11.73%±0.54%、23.97%±0.68%、10.45%±0.16%和8.59%±0.74%（P<0.05）。TUNEL法检测到HNE1细胞的晚期凋亡，各实验组增强了诱导凋亡作用（P<0.05）。唑来膦酸后用紫杉醇的治疗方案优于其他方案（P<0.05）。这种作用是由于在mRNA和蛋白质表达水平上抗凋亡蛋白Bcl-2下调，促凋亡蛋白Bax、Caspase3和Caspase9上调。唑来膦酸后用紫杉醇组的调节作用更大。
唑来膦酸可增强紫杉醇对HNE1细胞的体外抗增殖及诱导凋亡作用。唑来膦酸后用紫杉醇的治疗方案可能是最佳方案。凋亡的协同诱导是通过Bcl-2家族的作用并通过线粒体途径实现的。
