茉莉酸甲酯对人胰腺癌细胞株HS766T抑制作用的研究

[目的]观察茉莉酸甲酯对人胰腺癌细胞株HS766T的抑制增殖及诱导凋亡的作用。[方法]用0~10mmol/L浓度的茉莉酸甲酯体外处理人胰腺癌细胞株HS766T,分别在24h、48h、72h、96h用MTT法检测其细胞生长活性;PI单染色法检测其细胞周期;Annexin V/PI双染色法检测其细胞凋亡;RT-PCR、Western Blot检测基因Bcl-2、Bax及其蛋白的表达量。[结果]用0~10mmol/L茉莉酸甲酯处理胰腺癌细胞24h、48h、72h、96h后,细胞抑制率分别为:14.40%~71.41%、23.62%~89.68%、29.30%~94.76%、37.50%~94.74%;经0.001、0.01mmol/L的茉莉酸甲酯分别作用24h、48h后,与对照组相比,茉莉酸甲酯处理组处于G0/G1/S期的比例升高,G2/M期的比例降低;同时人胰腺癌细胞在0.001、0.01 mmol/L茉莉酸甲酯处理24h、48h后,对照组凋亡率为1.2%、1.6%,0.001mmol/L浓度组凋亡率为12.3%、16.8%,0.01mmol/L浓度组凋亡率为14.5%、23.2%;RTPCR检测显示Bax和Bcl-2在不同浓度和时间的扩增倍数分别为:1.18~1.48、0.92~0.33;Western Blot检测结果显示,Bax表达呈递增而Bcl-2表达呈递减的规律。[结论]不同浓度的茉莉酸甲酯对人胰腺癌细胞株HS766T细胞均有抑制作用,且呈明显的浓度依赖性,48h前细胞抑制率与药物浓度呈正相关,并可降低抑凋亡基因Bcl-2及升高促凋亡基因Bax的表达,从而抑制该细胞株体外生长和促进其凋亡。

Effect of methyl jasmonate on the proliferation and apoptosis induction in human pancreatic cancer cell line HS766T

[Objective]To investigate the effect of induction in human pancreatic cancer cell line HS766 monate in vitro on different concentration range with methyl jasmonate on the proliferation and apoptosis T. [Methods] HS766T was treated with methyl jas- 0-10 mmol/L and the cell growing activity was as- sayed by MTT at 24 h,48 h,72 h and 96 h,repectively. The cell cycle was detected by PI single staining, cell apoptosis was examed by Annexin V/PI double staining method,mRNA and protein expression of gene Bcl-2 and Bax were detected by RT-PCR and Western blot. I-Results]After treated with methyl jasmonate at 24 h,48 h,72 h and 96 h,cell inhibition rate were 14.40%-71.41%,23.62%-89. 68%,29.30%-94.76%, 37.50%-94.74% respectively. Comparing with control,the cell treated with methyl jasmonate after 24 h and 48 h at 0. 001 and 0.01 mmol/L increased the proportion during G0/G1/S phase transition and de- creased during G2/M phase as well. The apoptosis rate of cells treated with methyl jasmonate after 24 h and 48 h were 1.2%,1.6% in control group,12.3% and 16.8% at 0. 001 mmol/L,14.5% and 23.2% at 0.01 mmol/L. Accordingly,the mRNA level of gene Bax and Bcl-2 increased at different time point and differentconcentration with the proliferation rate 1.18-1.48 and 0.92-0.33 respectively. Moreover, the protein expression of Bax elevated as well as Bcl-2 reduced. [Conclusion3We demonstrated methyl jasmonate with different concentrations plays the inhibition role on HS766T in significant concentration-dependent man- ner. Within a definite period of time, cell inhibition rate correlates positively with the medicine concentra- tion. Furthermore,methyl jasmonate can decrease the expression of anti-apoptotic gene Bcl-2 as well as in- crease the expression of apoptotic gene Bax so as to inhibit the growth and promote apoptosis of human pancreatic cancer cell line HS766T.