| 引用本文:赵锦燕1，2，3，朱晓勤1，2，3，蓝紫林1，李斐1，林明和1，林久茂1，2，3.清解扶正颗粒促进凋亡和自噬逆转结直肠癌细胞多药耐药研究[J].世界中医药,2024,(15):. |
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| 清解扶正颗粒促进凋亡和自噬逆转结直肠癌细胞多药耐药研究 |
| Qingjie Fuzheng Granules Reverse Multidrug Resistance of Colorectal Cancer Resistant Cells via Inducing Apoptosis and Autophagy |
| 投稿时间：2024-01-18 |
| DOI：10.3969/j.issn.1673-7202.2024.15.005 |
| 中文关键词: 清解扶正颗粒 结直肠癌 多药耐药 细胞凋亡 细胞自噬 克隆形成 逆转 泵 |
| English Keywords:Qingjie Fuzheng Granules Colorectal cancer Multidrug resistance Apoptosis Autophagy Colony formation Reverse Pump |
| 基金项目:国家自然科学基金项目（81303125）；福建省自然科学基金项目（2021J01939，2022J01368） |
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| 中文摘要: |
| 目的：探讨清解扶正颗粒（QFG）对人结直肠癌HCT-8/5-FU耐药细胞的逆转作用机制。方法：培养人结直肠癌HCT-8细胞及HCT-8/5-FU耐药细胞，计算不同化疗药物的耐药指数（RI）和QFG的逆转倍数（RF）；设立空白对照组（0 mg/mL）和不同浓度（0.25、0.5、1.0 mg/mL）QFG干预组；各组干预48 h后，通过罗丹明-123（Rho123）染色和多柔比星（DOX）染色检测细胞外排泵功能，通过克隆形成试验检测细胞存活能力，应用磷脂酰丝氨酸（Annexin V-FITC）/碘化丙啶（PI）双染检测细胞凋亡，应用Cyto-ID染色检测细胞自噬，应用蛋白质印迹法检测P-糖蛋白（gP）、B细胞淋巴瘤-2(Bcl-2）、Bcl-2相关X蛋白（Bax）、选择性自噬接头蛋白（p62/SQSTM1）、微管相关蛋白1轻链（LC）3Ⅱ的蛋白表达。结果：HCT-8/5-FU细胞对5-氟尿嘧啶（5-FU）、多柔比星（DOX）和顺铂（DDP）均耐药，QFG可有效逆转化疗耐药，与空白对照组比较，不同浓度QFG显著增加Rho123、DOX和Cyto-ID的平均荧光强度（均P<0.05）；抑制细胞的克隆形成率（P<0.05），提高细胞凋亡率（P<0.05）；和空白对照组比较，QFG抑制P-gP、p62的蛋白表达（P<0.05）,降低Bcl-2/Bax比值（P<0.05），促进LC3-Ⅱ的蛋白表达（P<0.05）。结论：QFG促进细胞凋亡和自噬，抑制耐药细胞对化疗药物的外排泵功能，从而逆转多药耐药。 |
| English Summary: |
| To investigate the mechanism by which Qingjie Fuzheng Granules(QFG) reverse drug resistance in human colorectal cancer HCT-8/5-FU resistant cells.Methods:HCT-8 cells and HCT-8/5-FU resistant cells were cultured，and the resistance index(RI) for different chemotherapeutic drugs and the reversal fold(RF) of QFG were calculated.The cells were divided into a blank control group(0 mg/mL) and QFG intervention groups with different concentrations(0.25，0.5，1.0 mg/mL).After 48 hours of intervention，the efflux pump function was assessed by rhodamine-123(Rho123) and doxorubicin(DOX) staining.Cell survival was evaluated by colony formation assay.Apoptosis was detected using Annexin V-FITC/propidium iodide(PI) double staining.Autophagy was analyzed using Cyto-ID staining.Protein expression levels of P-glycoprotein(P-gP)，B-cell lymphoma 2(Bcl-2)，Bcl-2-associated X protein(Bax)，selective autophagy receptor protein(p62/SQSTM1)，and microtubule-associated protein 1 light chain 3 Ⅱ(LC3-Ⅱ) were measured by Western blot.Results:HCT-8/5-FU cells showed resistance to 5-fluorouracil(5-FU)，DOX，and cisplatin(DDP).QFG effectively reversed this drug resistance.Compared with the blank control group，QFG significantly increased the average fluorescence intensity of Rho123，DOX，and Cyto-ID(all P<0.05)，inhibited cell colony formation(P<0.05)，and increased the apoptosis rate(P<0.05).QFG also reduced the protein expression of P-gP and p62(P<0.05)，decreased the Bcl-2/Bax ratio(P<0.05)，and promoted the protein expression of LC3-Ⅱ(P<0.05).Conclusion:QFG can promote apoptosis and autophagy，inhibit the efflux pump function of resistant cells，and thereby reverse multidrug resistance. |
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