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目的研究夏枯草提取物(EPV)对人乳头瘤病毒(HPV)阳性人宫颈癌细胞的凋亡作用。方法培养人宫颈癌细胞Si Ha、He La(HPV阳性)及C-33-A(HPV阴性),与20、40、80μg/m L的EPV共孵育24 h,对照组加入等体积的DMSO,采用噻唑蓝(MTT)法检测细胞存活率;流式细胞术Annexin V/PI双染法检测细胞凋亡及细胞周期。取对数生长期的Si Ha、He La细胞,与40μg/m L EPV共孵育24 h,Western blotting法检测凋亡相关蛋白caspase 3、caspase 9、Bcl-2、Bax的表达情况。结果 EPV剂量相关地抑制HPV阳性人宫颈癌细胞Si Ha、He La活力(P<0.05、0.01、0.001);诱导Si Ha、He La细胞凋亡(P<0.05、0.01),使Si Ha、He La细胞周期阻滞在G0/G1期,抑制其增殖;对C-33-A细胞无显著作用。显著上调Si Ha、He La细胞内Cle caspase 3、Cle caspase 9、Bax表达,下调Bcl-2表达(P<0.05、0.01、0.001)。结论 EPV可诱导HPV阳性人宫颈癌Si Ha、He La细胞凋亡,对HPV阴性C-33-A细胞无显著作用,为夏枯草及其制剂在临床上治疗宫颈癌提供依据。
Objective To study the apoptosis effect of Prunella vulgaris extract (EPV) on human papillomavirus (HPV) positive human cervical carcinoma cells. Methods Cultured human cervical cancer cells Si Ha, He La (HPV positive) and C-33-A (HPV negative) were incubated with 20,40,80μg / ml EPV for 24 hours, Cell viability was detected by MTT assay. Cell apoptosis and cell cycle were detected by flow cytometry with Annexin V / PI double staining. Logarithmic growth phase Si Ha and He La cells were incubated with 40 μg / ml EPV for 24 h. The expressions of caspase 3, caspase 9, Bcl-2 and Bax were detected by Western blotting. Results EPV dose-dependently inhibited Si Ha and He La HPV-positive human cervical cancer cells (P <0.05, 0.01 and 0.001), induced apoptosis of Si Ha and He La cells (P <0.05 and 0.01) La cell cycle arrest in G0 / G1 phase, inhibition of its proliferation; C-33-A cells had no significant effect. Significantly upregulated the expressions of Cle caspase 3, Cle caspase 9 and Bax in Si Ha and He La cells, and down-regulated the expression of Bcl-2 (P <0.05,0.01,0.001). Conclusion EPV can induce apoptosis of SiHa and He La cells in HPV-positive human cervical cancer cells, and has no significant effect on HPV-negative C-33-A cells. It provides the basis for Prunella vulgaris and its preparation to treat cervical cancer clinically.