紫草素对感染HPV16的宫颈癌Caski细胞增殖与凋亡的影响

Effects of Shikonin on Cell Proliferation and Apoptosis of Cervical Cancer Caski Cells with HPV16

目的:研究紫草素(shikonin)对感染HPV16的宫颈癌Caski细胞作用。方法:对宫颈癌Caski细胞进行HPV病毒基因鉴定,将紫草素配成不同浓度(0、5、10、20、40、80μmol·L^(-1))作用于体外培养的宫颈癌Caski细胞,用MTT方法检测不同浓度紫草素作用不同时间(12 h、24 h、36 h)后对Caski细胞增殖能力的影响;采用Annexin V/PI试剂染色不同浓度紫草素(5、10、20、40μmol·L^(-1))作用24 h后的宫颈癌Caski细胞,上流式细胞仪观察细胞凋亡改变;用DAPI染色观察5、10、20、40μmol·L^(-1)紫草素作用于Caski细胞24 h后的凋亡形态学变化。结果:紫草素对宫颈癌Caski细胞具有抑制增殖、促进凋亡作用,在一定范围内,随着浓度和时间增加,抑制增殖作用逐渐增加;凋亡数目逐渐增多,其中20、40μmol·L^(-1)的紫草素可明显抑制细胞增殖并诱导其凋亡,DAPI染色可观察到典型的细胞凋亡改变:细胞核碎裂、固缩;且细胞凋亡现象随紫草素浓度增加而愈加明显。结论:紫草素可抑制感染HPV16的宫颈癌Caski细胞增殖并促进其凋亡。

Objective: To investigate the effect of shikonin on proliferation and apoptosis of human cervical cancer Caski cells with HPV16.Methods: Cervical cancer Caski cells were identified by HPV virus gene. Then the human cervical cancer Caski cells were cultivated and divided into control group and shikonin group(the concentration was respectively 5,10,20,40 and 80 μmol·L-1). The effect of shikonin on cell viability of cells cultivated were studied by MTT assay(12 h, 24 h and 36 h). Then Shikonin respectively 5,10,20,40 μmol·L-1 on apoptosis of cells cultivated after 24 h were studied by flow cytometric. Last Shikonin respectively 5,10,20,40 μmol·L-1 on apoptotic morphology of cells cultivated after 24 h were studied by DAPI staining. Results: The proliferation of Caski cells in cervical carcinoma shows inhibitory effect on the proliferation of cervical cancer and the concentration and time dependence. Apoptosis rate increased with the increase of concentration of shikonin, 20 and 40 μmol·L-1 of shikonin can obviously inhibit cell proliferation and induce its apoptosis. Typical apoptosis morphological changes observing by DAPI staining, including disintegrate and pyknotic nucleus, are more and more obvious following the increase of the concentration. Conclusion: Shikonin can inhibit the proliferation of cervical cancer Caski cells with HPV16 and promote the apoptosis of cancer cells.