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北京中医药大学 中药学院 中药现代研究中心,北京 100029
黄惠铭,在读硕士,从事中药活性成分抗肿瘤作用研究,E-mail:hhmtcm@163.com
胡仲冬,研究员,从事中药活性成分抗肿瘤作用研究,Tel:010-64286180,E-mail:huzhongdong@126.com
收稿日期:2021-06-24,
网络出版日期:2021-09-08,
纸质出版日期:2022-01-05
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黄惠铭,田颖颖,庞道然等.血竭石油醚提取物对人胃癌HGC-27和MGC-803细胞的抗肿瘤作用[J].中国实验方剂学杂志,2022,28(01):85-91.
HUANG Hui-ming,TIAN Ying-ying,PANG Dao-ran,et al.Anti-tumor Effect of Draconis SanguisPetroleum Ether Fraction on Human Gastric Cancer HGC-27 and MGC-803 Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(01):85-91.
黄惠铭,田颖颖,庞道然等.血竭石油醚提取物对人胃癌HGC-27和MGC-803细胞的抗肿瘤作用[J].中国实验方剂学杂志,2022,28(01):85-91. DOI: 10.13422/j.cnki.syfjx.20212124.
HUANG Hui-ming,TIAN Ying-ying,PANG Dao-ran,et al.Anti-tumor Effect of Draconis SanguisPetroleum Ether Fraction on Human Gastric Cancer HGC-27 and MGC-803 Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(01):85-91. DOI: 10.13422/j.cnki.syfjx.20212124.
目的
2
研究血竭石油醚提取物(SDPEF)对人胃癌HGC-27和MGC-803细胞增殖、凋亡、体外迁移能力及自噬的影响,并阐明其分子作用机制。
方法
2
采用细胞增殖与活性检测(CCK-8)法检测SDPEF(0, 20, 40, 60, 80 mg·L
-1
)作用24,48,72 h时,对人胃癌HGC-27和MGC-803细胞体外增殖的影响。借助Hoechst染色实验观察不同浓度的SDPEF作用48 h后对HGC-27和MGC-803细胞凋亡的影响。采用流式细胞术检测不同浓度SDPEF作用48 h,对HGC-27和MGC-803细胞凋亡率的影响。采用细胞划痕实验观察不同浓度SDPEF在不同作用时间点对人胃癌HGC-27和MGC-803细胞体外迁移能力的影响。借助吖啶橙染色实验检测不同浓度SDPEF对HGC-27和MGC-803细胞自噬的影响。利用蛋白免疫印迹法检测SDPEF作用人胃癌HGC-27和MGC-803细胞48 h后对信号通路相关蛋白表达水平的调控作用。
结果
2
与空白组比较,SDPEF(30 mg·L
-1
)组人胃癌HGC-27和MGC-803细胞的增殖和体外迁移能力明显降低(
P<
0.05),且具有浓度与时间依赖性;SDPEF(60 mg·L
-1
)还能够诱导人胃癌HGC-27和MGC-803细胞凋亡(
P<
0.01)和自噬的发生。与空白组比较,SDPEF(60 mg·L
-1
)能够下调磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)表达(
P<
0.05,
P<
0.01),且能够下调磷酸化信号传导与转录激活因子3(p-STAT3)蛋白表达水平(
P<
0.01),提示SDPEF可能是通过抑制mTOR/STAT3信号通路来抑制人胃癌HGC-27和MGC-803细胞的增殖、迁移并诱导其发生凋亡和自噬。
结论
2
mTOR/STAT3信号通路下调可能参与了SDPEF的抗胃癌作用。该研究能够为血竭抗肿瘤作用研究提供一定的参考。
Objective
2
To investigate the effect of Draconis Sanguis petroleum ether fraction (DSPEF) on the proliferation, apoptosis, migration, and autophagy of human gastric cancer HGC-27 and MGC-803 cells, and preliminarily elucidate its molecular mechanism.
Method
2
Cell counting kit-8 (CCK-8) assay was used to detect the effect of DSPEF at different concentrations (0, 20, 40, 60, 80 mg·L
-1
) on the proliferation of HGC-27 and MGC-803 cells after 24, 48, 72 h. Hoechst staining and flow cytometry were used to explore the effects of DSPEF at different concentrations on the apoptosis and apoptosis rate of HGC-27 and MGC-803 cells after 48 h treatment, respectively. The wound healing assay and acridine orange staining were used to investigate the effects of DSPEF on the migration and autophagy of HGC-27 and MGC-803 cells, respectively. Western blot was used to detect the expression levels of signaling pathway-related proteins in HGC-27 and MGC-803 cells treated with DSPEF for 48 h.
Result
2
Compared with the control group, DSPEF(30 mg·L
-1
) inhibited the proliferation and migration of HGC-27 and MGC-803 cells in a concentration- and time-dependent manner (
P
<
0.05), and induced the apoptosis (
P
<
0.01) and autophagy of HGC-27 and MGC-803 cells. DSPEF (60 mg·L
-1
) down-regulated the protein levels of phosphorylated mammalian target of rapamycin (p-mTOR) (
P<
0.05,
P<
0.01) and down-regulated phospho-signal transducer and activator of transcription 3 (p-STAT3) in HGC-27 and MGC-803 cells (
P
<
0.01), suggesting that DSPEF presumedly inhibited the proliferation and migration of human gastric cancer HGC-27 and MGC-803 cells and induced their apoptosis and autophagy by inhibiting the mTOR/STAT3 signaling pathway.
Conclusion
2
The down-regulation of the mTOR/STAT3 signaling pathway may be involved in the anti-gastric cancer effect of DSPEF. This study is expected to provide a reference for the investigation of the anti-tumor effect of Draconis Sanguis
.
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