| 高爱春.复方苦参注射液通过增强细胞免疫杀伤作用诱导宫颈癌海拉细胞凋亡[J].安徽医药,2022,26(9):1724-1728. |
| 复方苦参注射液通过增强细胞免疫杀伤作用诱导宫颈癌海拉细胞凋亡 |
| Compound Kushen injection induces apoptosis of cervical cancer HeLa cells by enhancing cellular immune killing |
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| DOI：10.3969/j.issn.1009-6469.2022.09.007 |
| 中文关键词: HeLa细胞 中草药 复方苦参注射液 HLA抗原 细胞凋亡 |
| 英文关键词: HeLa cells Drugs, Chinese herbal Compound Kushen injection HLA antigens Apoptosis |
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| 中文摘要: |
| 目的探讨复方苦参注射液对人宫颈癌海拉细胞凋亡及人白细胞抗原G(HLA-G)基因表达的影响。方法2018年7月至2019年8月，体外培养人宫颈癌海拉细胞，用0、100、200、500 mL/L复方苦参注射液分别干预海拉细胞48 h。采用人胆囊收缩素/缩胆囊素八肽(CCK-8)法和平板克隆形成实验检测细胞增殖情况，流式细胞术检测细胞周期和凋亡情况，蛋白质印迹法检测细胞中B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、活化胱天蛋白酶-3(cleaved-caspase-3)蛋白水平，酶联免疫吸附测定(ELISA)检测细胞上清液中转化生长因子β1(TGF-β1)、肿瘤坏死因子-α(TNF-α)和白细胞介素-4(IL-4)含量，实时荧光定量逆转录聚合酶链反应(qRT-PCR)和蛋白质印迹法检测细胞中HLA-G表达水平。结果与0 mL/L复方苦参注射液干预组相比，经100、200、500 mL/L复方苦参注射液干预的海拉细胞增殖能力明显减弱，G0/G1期细胞比例明显增多，S期细胞比例明显减少，细胞凋亡率［(6.02±0.35)%比(14.38±1.06)%、(20.62±1.57)%、(31.84±2.11)%］升高，Bax和cleaved-caspase-3蛋白水平明显上调，Bcl-2蛋白水平明显下调，细胞上清液中TGF-β1和IL-4含量明显降低，TNF-α含量明显升高，HLA-G mRNA和蛋白表达下降，且其作用具有剂量依赖关系(P<0.05)。结论复方苦参注射液能够诱导人宫颈癌海拉细胞凋亡，其作用机制可能与抑制HLA-G基因的表达来增强细胞免疫杀伤作用有关。 |
| 英文摘要: |
| Objective To investigate the effect of compound Kushen injection on apoptosis of human cervical carcinoma HeLa cells and expression of human leukocyte antigen G (HLA-G) gene.Methods The starting and ending time of this experiment was from July 2018 to August 2019, Human cervical carcinoma HeLa cells were cultured in vitro. HeLa cells were treated with 0, 100, 200 and 500mL/L compound Kushen injection for 48 h. Cell proliferation was detected by cell counting kit (CCK-8) method and plate colony formation assay. Cell cycle and apoptosis were detected by flow cytometry. Western blotting was used to detect Bcl-2-related X protein (Bax),B-cell lymphoma/leukemia-2 (Bcl-2), activated cysteine-containing aspartate proteolytic enzyme 3 (cleaved-caspase-3) protein level; enzyme-linked immunosorbent assay (ELISA) to detect the contents of transforming growth factor β1 (TGF-β1), tumor necrosis factor-α(TNF- α) and interleukin-4 (IL-4); HLA-G expression level in cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting.Results Compared with the 0 mL/L compound Kushen injection group, the proliferation ability of HeLa cells intervened by 100, 200 and 500 mL/L compound Kushen injection were significantly attenuated, the proportion of cells in G0/G1 phase was significantly increased, the proportion of cells in S phase was significantly reduced, and the apoptosis rate [(6.02±0.35)% h vs. (14.38±1.06)%, (20.62±1.57)%, (31.84±2.11)%], Bax and cleaved-caspase-3 protein levels were significantly up-regulated,Bcl-2 protein levels were significantly down-regulated, TGF- β1 and IL-4 levels were significantly decreased in cell supernatants,TNF-α levels were significantly increased, HLA-G mRNA and protein expression were significantly decreased, and its effect was dose dependent (P<0.05).Conclusion Compound Kushen injection can induce apoptosis of human cervical cancer HeLa cells, and its mechanism may be related to inhibition of HLA-G gene expression to enhance the killing of cellular immune. |
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