Hedgehog通路对肺腺癌增殖、凋亡及间隙连接蛋白Cx32和Cx43表达的影响

EFFECT OF THE HEDGEHOG PATHWAY ON THE PROLIFERATION AND APOPTOSIS OF LUNG ADENOCARCINOMA CELLS AND THE EXPRESSION OF CONNEXINS CX32 AND CX43 IN THESE CELLS

目的检测阻断Hedgehog通路对肺腺癌细胞增殖、凋亡及间隙连接蛋白Cx32和Cx43表达的影响,初步探索其在肺腺癌增殖和转移中的作用机制。方法取对数生长期肺腺癌细胞A549,分为阴性对照组及环巴胺处理组,分别以0、10、20、30和40μmol/L环巴胺处理A549细胞24、48和72 h后,应用MTT法测定其生长抑制率;以40μmol/L环巴胺处理A549细胞48 h后,以Annexin V/PI双染流式细胞术检测细胞凋亡率及Cx32、Cx43胞膜阳性表达率。结果环巴胺处理后,A549细胞增殖率明显下降,且表现为浓度、时间依赖性;40μmol/L环巴胺作用于A549细胞48 h后,细胞凋亡率明显高于空白对照组(P<0.05);胞膜Cx32和Cx43阳性表达率均明显高于空白对照组(P<0.05)。结论 Hedgehog通路可能抑制肺腺癌细胞凋亡,并调节间隙连接蛋白Cx43及Cx32胞膜的表达。

Objective To investigate the effect of the Hedgehog pathway on the proliferation and apoptosis of lung adenocarcinoma cells and the expression of connexins Cx32 and Cx43 in these cells,as well as the mechanism of action of the Hedgehog pathway in the proliferation and metastasis of lung adenocarcinoma. Methods Lung adenocarcinoma A549 cells in the logarithmic phase were selected and divided into negative control group and cyclopamine treatment group. After A549 cells were treated with 0（ negative control）,10,20,30,and 40 μmol/L cyclopamine for 24,48,and 72 h,MTT assay was used to measure growth inhibition rate. After A549 cells were treated with40 μmol/L cyclopamine for 48 h,Annexin V/PI double-staining flow cytometry was used to measure cell apoptosis rate and positive rates of Cx32 and Cx43 in the membrane. Results After cyclopamine treatment,the proliferation rate of A549 cells decreased significantly in a dose-and time-dependent manner. Compared with the blank control group,the group of A549 cells treated with 40 μmol/L cyclopamine for48 h had significantly higher cell apoptosis rate（ P〈0.05） and positive rates of Cx32 and Cx43 in the membrane（ P〈0.05）. Conclusion The Hedgehog pathway can inhibit the apoptosis of lung adenocarcinoma cells and regulate the expression of connexins Cx32 and Cx43.