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左旋紫草素对顺铂耐药人宫颈癌HeLa细胞的逆转作用研究
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| 篇名： | 左旋紫草素对顺铂耐药人宫颈癌HeLa细胞的逆转作用研究 |
| TITLE： | Study on Reversal Effects of Levoshikonin on Cisplatin Resistance of Human Cervical Carcinoma HeLa Cells |
| 摘要： | 目的：探讨左旋紫草素（L-SHK）对顺铂（DDP）耐药人宫颈癌HeLa细胞（HeLa/DDP）的逆转作用及其可能机制。方法：以人宫颈癌HeLa细胞株为研究对象，以DDP诱导获得HeLa/DDP耐药细胞；采用CCK-8法测定HeLa/DDP细胞的耐药指数以及不同剂量L-SHK（0.125、0.25、0.5、1、2、4、8、16μmol/L）对该细胞的抑制率、半数抑制浓度（IC50）和逆转倍数；采用流式细胞术检测低、中、高剂量L-SHK（0.3、0.6、1.2μmol/L）联合DDP对HeLa/DDP细胞周期及凋亡率的影响，采用Westernblotting法检测低、中、高剂量L-SHK（0.3、0.6、1.2μmol/L）联合DDP对HeLa/DDP细胞凋亡相关蛋白[剪切型胱天蛋白酶3（Cleavedcaspase-3）、Bcl-2、Bax]表达的影响。结果：所得HeLa/DDP细胞的耐药指数为11.8。L-SHK对HeLa/DDP细胞的抑制率有随剂量增加而逐渐升高的趋势。与单用DDP比较，DDP+低、中、高剂量L-SHK组细胞的IC50值均显著降低，且呈剂量依赖性（P＜0.05）；DDP+低、中、高剂量L-SHK组的逆转倍数分别为1.38、2.80、6.71。与空白对照组比较，各药物组G0/G1期、S期细胞比例，各剂量L-SHK联用组细胞的早期、晚期凋亡率及总凋亡率以及Bax、Cleavedcaspase-3蛋白的表达量均显著升高；各药物组G2/M期细胞比例以及各剂量L-SHK联用组Bcl-2蛋白的表达量均显著降低（P＜0.05）。与DDP组比较，各剂量L-SHK联用组S期、G2/M期细胞比例和Bcl-2蛋白的表达量均显著降低；G0/G1期细胞比例，早期、晚期凋亡率及总凋亡率和Bax、Cleavedcaspase-3蛋白的表达量均显著升高（P＜0.05）。结论：HeLa/DDP细胞对DDP具有一定的耐药性，L-SHK可逆转这种耐药性。L-SHK和DDP联用可促进HeLa/DDP细胞凋亡，且作用强于DDP单用；这种作用可能与影响细胞周期、调控凋亡相关蛋白的表达有关。 |
| ABSTRACT： | OBJECTIVE：To investigate the reversal effects and potential mechanism of levoshikonin （L-SHK）on cisplatin （DDP）resistance of human cervical carcinoma HeLa cells. METHODS ：Human cervical carcinoma HeLa cells were used as research objects ，and drug-resistant HeLa/DDP cells were induced by DDP. CCK- 8 assay was used to determine drug resistance index of HeLa/DDP cells ，the inhibition rate of different doses of L-SHK （0.125，0.25，0.5，1，2，4，8，16 μmol/L）on cell proliferation，IC50 and the reversion index of L-SHK on HeLa/DDP cells. Effects of low ，medium and high doses of L-SHK （0.3， 0.6，1.2 μmol/L）combined with DDP on cell cycle and apoptosis rate were determined by flow cytometry. Western blotting assay was used to detect the effects of low ，medium and high doses of L-SHK （0.3，0.6，1.2 μmol/L）combined with DDP on the expression of apoptosis-related protein （Cleaved caspase- 3，Bcl-2 and Bax ）. RESULTS ：The drug resistance index of HeLa/DDP cells was 11.8. The inhibition rate of L-SHK on HeLa/DDP cells increased with the increase of dose. Compared with DDP alone ， IC50 of DDP+low-dose ，medium-dose and high-dose L-SHK groups were decreased significantly ，with a dose-dependent manner （P＜0.05）. The reversion indexes were 1.38，2.80，6.71 in DDP+low-dose ，medium-dose and high-dose L-SHK groups. Compared with blank control group ，the proportion of cells at phase G 0/G1 and phase S in administration groups ，as well as early and late apoptosis rate and total apoptosis rate of cells ，the protein expression of Bax and Cleaved caspase- 3 in L-SHK combination groups were increased significantly ；the proportion of cells at phase G 2/M in administration group as well as the protein expression of Bcl-2 in L-SHK combination groups were decreased significantly （P＜0.05）. Compared with DDP group ，the proportion of cells at phase S and G 2/M and the protein expression of Bcl- 2 in L-SHK combination groups were significantly decreased ；the proportion of cells at phase G 0/G1，early and late apoptosis rate and total apoptosis rate ，the protein expression of Bax and Cleaved caspase- 3 in L-SHK combination groups were significantly increased （P＜0.05）. CONCLUSIONS ：HeLa/DDP cells are resistant to DDP ，and L-SHK can reverse the drug resistance. L-SHK combined with DDP can promote the apoptosis of HeLa/DDP cells ，which is better than DDP alone. Its mechanism may be related to the influence of cell cycle and the regulation of apoptosis-related protein expression. |
| 期刊： | 2020年第31卷第15期 |
| 作者： | 杜春双，马亚妮，王帅，张飞，张洁，桑广健 |
| AUTHORS： | DU Chunshuang，MA Yani，WANG Shuai，ZHANG Fei，ZHANG Jie，SANG Guangjian |
| 关键字： | 人宫颈癌HeLa细胞；左旋紫草素；顺铂；耐药；凋亡；细胞周期；凋亡蛋白；逆转作用；机制 |
| KEYWORDS： | Human cervical carcinoma HeLa cells ；Levoshikonin；Cisplatin；Resistance；Apoptosis；Cell cycle ；Apoptosis- |
| 阅读数： | 417 次 |
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