目的 研究夏枯草提取物(EPV)對人乳頭瘤病毒(HPV)陽性人宮頸癌細(xì)胞的凋亡作用。方法 培養(yǎng)人宮頸癌細(xì)胞SiHa、HeLa(HPV陽性)及C-33-A(HPV陰性),與20、40、80 μg/mL的EPV共孵育24 h,對照組加入等體積的DMSO,采用噻唑藍(lán)(MTT)法檢測細(xì)胞存活率;流式細(xì)胞術(shù)Annexin V/PI雙染法檢測細(xì)胞凋亡及細(xì)胞周期。取對數(shù)生長期的SiHa、HeLa細(xì)胞,與40 μg/mL EPV共孵育24 h,Western blotting法檢測凋亡相關(guān)蛋白caspase 3、caspase 9、Bcl-2、Bax的表達(dá)情況。結(jié)果 EPV劑量相關(guān)地抑制HPV陽性人宮頸癌細(xì)胞SiHa、HeLa活力(P<0.05、0.01、0.001);誘導(dǎo)SiHa、HeLa細(xì)胞凋亡(P<0.05、0.01),使SiHa、HeLa細(xì)胞周期阻滯在G₀/G₁期,抑制其增殖;對C-33-A細(xì)胞無顯著作用。顯著上調(diào)SiHa、HeLa細(xì)胞內(nèi)Cle caspase 3、Cle caspase 9、Bax表達(dá),下調(diào)Bcl-2表達(dá)(P<0.05、0.01、0.001)。結(jié)論 EPV可誘導(dǎo)HPV陽性人宮頸癌SiHa、HeLa細(xì)胞凋亡,對HPV陰性C-33-A細(xì)胞無顯著作用,為夏枯草及其制劑在臨床上治療宮頸癌提供依據(jù)。

Objective To study the potential apoptotic effect of extract from Prunella vulgaris (EPV), a traditional Chinese medicine, on human papilloma virus (HPV) positive and negative human cervical cancer cells. Methods Human cervical carcinoma cell lines SiHa, Hela (HPV positive), and C-33-A (HPV negative) were cultured, and treated with 20, 40, and 80 μg/mL of EPV for 24 h. Control group were treated with equal volume of DMSO. The survival rate of cells was detected by methyl thiazolyl tetrazolium (MTT) method. Flow cytometry Annexin V/PI double staining method was used to detect cell apoptosis and cell cycle. SiHa and HeLa cells were incubated with 40 μg/mL EPV for 24 h, and Western blotting was used to detect the expression of apoptosis related protein caspase 3, caspase 9, Bcl-2, and Bax. Results EPV reduced the viability of SiHa and HeLa cells dose-dependently (P<0.05, 0.01 and 0.001). Moreover, EPV induced apoptosis in SiHa and HeLa cells (P<0.05 and 0.01), inhibited the proliferation of SiHa and HeLa cells through arresting their cycle in G₀/G₁ phase. However, EPV did not show significantly effect on C-33-A cells. Western blotting results showed that Cle caspase 3, Cle caspase 9, Bax, and Bcl-2 proteins were involved in EPV induced cell apoptosis (P<0.05, 0.01, and 0.001). Conclusion EPV could induce apoptosis in HPV-positive cervical cancer cells, which provides novel information for clinical application of P. vulgaris and its preparations for the HPV-positive cervical cancer patients.