目的：通过构建基因真核表达载体,探讨人2型大麻素受体(human cannabinoid receptor 2，hCB2R)对人子宫颈癌HeLa细胞体外凋亡的作用及机制。方法：选用人脑组织的cDNA作为模板,进行hCB2R基因的RT-PCR扩增,构建重组质粒GV230-hCB2R及其对照空质粒GV230并转染HeLa细胞，Western blotting法及免疫荧光细胞化学染色联合激光扫描共聚焦显微镜技术检测hCB2R表达及细胞内定位；流式细胞术检测HeLa细胞凋亡，Western blotting法及实时荧光定量PCR检测HeLa细胞中hCB2R、Bcl-2、Bax、Bad的表达。结果：与空质粒转染组相比，GV230-hCB2R转染HeLa细胞后表达相对分子质量40 000的hCB2R蛋白，且细胞膜和细胞质中均有hCB2R的表达；GV230-hCB2R转染组的细胞凋亡率显著高于GV230空质粒对照组\[(14.51±4.51)% vs（6.29±0.57）%，t=1.72, P<0.05\]；与空质粒对照组相比，hCB2R转染组细胞内Bax和Bad的表达水平明显上调（P＜0.05）, 而Bcl-2的表达明显下调（P<0.05）。结论： hCB2R对子宫颈癌HeLa细胞的生长表现出明显的抑制作用，其作用机制可能与hCB2R直接参与了细胞凋亡相关蛋白的表达变化有关。

Objective:To construct a eukaryotic expression vector containing human cannabinoid receptor 2 (hCB2R) gene and investigate its effect on the apoptosis of cervical cancer HeLa cells as well as the possible mechanism. Methods：cDNA of human brain tissues was selected as template for the amplification of hCB2R gene by RT-PCR; recombinant plasmid GV230-hCB2R was constructed to transfect HeLa cells, and the empty vector GV230 was used as control vector. Western blotting and immunofluorescence staining combined with laser scanning and co-confocal microscope technology were used to detect hCB2R expression and intra-cecullar localization; Flow cytometry was used to determine the apoptosis of HeLa cells; Western blotting and real-time fluorescence quantitative PCR were used for the detection of expression of hCB2R, Bcl-2, Bax and Bad in HeLa cells. Results: Compared with empty plasmid group, GV230-hCB2R transfected HeLa cells expressed hCB2R protein with relative molecular weight (Mr) 40 000, on both cell membrane and cytoplasm; cell apoptosis rate of GV230-hCB2R group was significantly higher than that of the GV230 empty plasmid group (\[14.51±4.51\]% vs \[6.29±0.57\]%, t=1.72, P<0.05). Compared with the control group, the expression levels of Bax and Bad were significantly increased (P<0.05) while the expression of Bcl-2 was significantly decreased (P<0.05) in GV230-hCB2R group. Conclusion:hCB2R has a significant inhibitory effect on the growth of cervical cancer HeLa cell line, and its mechanism may be directly related to the involvement of hCB2R in the expression of apoptosis related proteins.

重庆市科委前沿与基础研究资助项目（No. cstc2014jcyjA1215）；重庆市卫生局医学科研项目(No. 2012-1-096)