康莱特诱导人胰腺癌细胞凋亡的实验研究

目的 探讨康莱特(KLT)对人胰腺癌细胞凋亡的影响及其作用机制。方法 采用MTT法观察KLT对人胰腺癌细胞株8988细胞增殖的抑制作用。采用TUNEL染色、DNA梯度电泳法和流式细胞术检测细胞凋亡改变，并以RT-PCR和Western blot检测凋亡调节基因p53和bcl-2的表达。结果 KLT能明显抑制人胰腺癌8988细胞的生长，且呈时间和浓度依赖性。KLT作用后8988细胞呈现凋亡特征，TUNEL染色可见发黄绿色荧光的凋亡细胞，DNA电泳可见典型梯形条带，流式细胞术显示凋亡细胞比例升高。RT-PCR和Western blot检测可见p53基因表达显著增加，而bcl-2基因表达减少。结论 KLT能诱导人胰腺癌细胞凋亡，其作用可能与凋亡调节基因p53的上调和bcl-2的下调有关。

Experimental study on the effect of Kanglaite on apoptosis of human pancreatic adenocarcinoma cells

Objective To investigate the effect of Kanglaite(KLT) on human pancreatic adenocarcinoma cells and its possible mechanism.Methods Growth inhibition was measured by methyl thiazolyl tetrazolium (MTT) assay. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method, flow cytometry and DNA fragmentation assay. The expression of apoptosis-regulated gene p53 and bcl-2 was analyzed by RT-PCR and Western blot.Results KLT inhibited the growth of human pancreatic adenocarcinoma cell line 8988 in a dose- and time-dependent manner. The cells treated with KLT had the characteristics of apoptosis, demonstrating as yellow-greenish fluorescence shown by TUNEL, typical ladder pattern of DNA fragments in DNA electrophoresis and increment of apoptotic proportion with flow cytometry. RT-PCR and Western blot showed that the expression of p53 gene was apparently increased by KLT, whereas the expression of bcl-2 gene was reduced.Conclusion KLT can induce apoptosis of human pancreatic adenocarcinoma cells in vitro, which may be mediated by up-regulating the apoptosis-regulated gene p53 and down-regulating the apoptosis-regulated gene bcl-2.