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Archive > Volume 23 Issue 5 > 2016,23(5):651-655. DOI:10.3872/j.issn.1007-385X.2016.05.011 Prev Next
Basic Research
Up-regulation of human cannabinoid receptor 2 induces apoptosis of cervical cancer HeLa cells
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Abstract:
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.
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Project Supported:
Project supported by the Frontier and Basic Research of Chongqing（No.cstc2014jcyjA1215），and the Medical Scientific Research of Chongqing Municipal Health Bureau（No.2012-1-096）
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