杜春双,王帅,张飞,于明欣,宋晓坤,王晨
DU Chunshuang,WANG Shuai,ZHANG Fei,YU Mingxin,SONG Xiaokun,WANG Chen

• 1:天津医科大学肿瘤医院国家肿瘤临床医学研究中心天津市"肿瘤防治"重点实验室
• 2:岳阳市广济医院

摘要(Abstract)：

目的探讨左旋紫草素(shikonin,SHK)联合顺铂(cisplatin,CDDP)对宫颈癌Hela细胞的抑制作用及作用机制。方法采用CCK-8法分别测定不同浓度的顺铂、紫草素和紫草素联合顺铂溶液对宫颈癌Hela细胞增殖的活性抑制作用;流式细胞仪测定紫草素联合顺铂后诱导宫颈癌Hela细胞凋亡的情况;Western blot检测Caspase-3蛋白表达情况。结果左旋紫草素对宫颈癌Hela细胞有抑制作用,24 h与48 h的IC50分别为1.05和0.89 mg·L~(-1)。紫草素联合顺铂作用于Hela细胞48 h后,细胞数量明显变少,胞体变圆,缩小,出现明显的细胞凋亡现象;不同浓度的紫草素联合顺铂作用于Hela细胞,24 h/48 h的IC50值(0.15 mg·L~(-1)SHK+CDDP 2.82 mg·L~(-1)/1.87 mg·L~(-1);0.3 mg·L~(-1)SHK+CDDP 2.43 mg·L~(-1)/1.52 mg·L~(-1);0.6 mg·L~(-1)SHK+CDDP 2.01 mg·L~(-1)/1.23 mg·L~(-1))明显低于顺铂(CDDP 3.54 mg·L~(-1)/2.78 mg·L~(-1))的IC50值,具有显著性差异(P<0.05);用流式细胞仪检测细胞凋亡率显示,随着紫草素浓度的增大,紫草素联合顺铂(1.2 mg·L~(-1)SHK+CDDP总凋亡率15.6%)细胞凋亡比率增高,与对照组顺铂(CDDP总凋亡率3.7%)相比,具有显著性差异(P<0.05)。0.3、0.6、1.2 mg·L~(-1)紫草素+顺铂分别作用Hela细胞48 h后,Western blot结果显示,Caspase-3随着紫草素浓度的增加,出现了明显的裂解活化,促进了凋亡。结论紫草素联合顺铂对宫颈癌Hela细胞有明显的抑制作用,其抑制作用与促进Hela细胞凋亡有关。
Objective To explore the inhibition mechanism of shikonin(SHK) combined with cisplatin(CDDP) on the proliferative activity of cervical cancer Hela cells. Methods A cell counting kit-8(CCK-8)was used to detect the inhibitory roles of CDDP and SHK as well as CDDP combined with different concentrations of SHK on the proliferation of cervical cancer Hela cells. Flowcytometer was used to detect the apoptosis of Hela cells induced by SHK with CDDP,and Western blot was conducted to detect the expression of caspase-3 protein. Results There is inhibitory effect on cervical cancer He La cells by SHK. The SHK combined with CDDP considerably reduced the cell viability of Hela cells after 48 h. In addition,the cell bodies of the Hela cells shrunk and became round,indicating the extensive apoptosis. Moreover,the IC50 values of the medications containing CDDP and different concentrations of SHK(the IC50 values of0. 15 mg·L~(-1) SHK + CDDP,0. 3 mg·L~(-1) SHK + CDDP and 0. 6 mg·L~(-1) SHK + CDDP were 2. 82 mg·L~(-1),2. 43 mg·L~(-1) and 2. 01 mg·L~(-1) at 24 h,respectively,and 1. 87 mg·L~(-1),1. 52 mg·L~(-1) and 1. 23 mg·L~(-1) at48 h,respectively) were considerably lower than that of the medication containing only CDDP(the IC50 values were 3. 54 mg·L~(-1) and 2. 78 mg·L~(-1) at 24 h and 48 h,respectively)(P < 0. 05). The flowcytometer results showed that the total apoptotic rate of the Hela cells administered with SHK combined with CDDP increased(15. 6% at 1. 2 mg·L~(-1) SHK + CDDP) and exhibited a significant difference(P < 0. 05) at the increasing concentration of SHK compared with that of the control group(3. 7%). Meanwhile,the results of Western blot analysis showed that the caspase-3 proteins were cleaved after the medications containing CDDP with SHK concentrations of 0. 3,0. 6 and 1. 2 mg·L~(-1) acted on the Hela cells for 48 h. The cleaving of the caspase-3 proteins promoted the apoptosis of the Hela cells. Conclusions SHK combined with CDDP has a significant inhibitory effect on cervical cancer Hela cells,and the inhibitory effect is related to its ability to induce the apoptosis of Hela cells.

关键词(KeyWords)： 左旋紫草素;顺铂;宫颈癌;Hela细胞
shikonin;cisplatin;cervical cancer;Hela cells

Abstract:

Keywords:

基金项目(Foundation): 天津市卫计委科技基金资助课题(2015KZ087)

作者(Author): 杜春双,王帅,张飞,于明欣,宋晓坤,王晨
DU Chunshuang,WANG Shuai,ZHANG Fei,YU Mingxin,SONG Xiaokun,WANG Chen

DOI: 10.14066/j.cnki.cn21-1349/r.2018.03.012

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