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张冬梅,张雅明.苦参碱逆转人结肠癌细胞株(HT-29)奥沙利铂耐药性的作用及机制研究[J].湖南中医药大学学报英文版,2016,36(11):22-26.[Click to copy
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| 苦参碱逆转人结肠癌细胞株(HT-29)奥沙利铂耐药性的作用及机制研究 |
| 张冬梅,张雅明 |
| (上海中医药大学附属曙光医院, 上海 200000) |
| 摘要: |
| 目的 研究苦参碱对结肠癌耐药细胞HT-29/奥沙利铂（Oxaliplatin,OXA）耐药性的逆转作用并探讨其可能的作用机制。方法 采用逐步增加药物浓度的方法建立奥沙利铂耐药结肠癌细胞株HT-29/OXA；CCK-8法测定苦参碱对HT-29/OXA细胞的耐药性逆转作用，流式细胞术检测细胞凋亡、周期变化；实时定量PCR检测各组细胞肺耐药蛋白（lung resistance protein LRP）和P-glycoprotein（P-gp）mRNA表达水平；Western blot检测各组细胞LRP和P-gp蛋白的表达水平。结果 苦参碱使HT-29/OXA细胞对奥沙利铂的敏感性增加，耐药性得到部分逆转。奥沙利铂（0.5 μg/mL）和苦参碱（3.0 μg/mL）单独用药对结肠癌耐药细胞株HT-29/OXA细胞周期以及细胞凋亡没有影响；联合用药对HT-29/OXA生长增殖具有明显抑制作用并且能够通过改变细胞周期引起凋亡（P<0.05）。奥沙利铂（0.5 μg/mL）和苦参碱（3.0 μg/mL）单独用药对HT-29/OXALRP和P-gp mRNA以及蛋白的表达没有影响；联合用药作用后LRP mRNA和P-gp mRNA表达水平降低（P<0.01），同时下调了LRP和P-gp蛋白表达（P<0.01）。结论 苦参碱部分逆转HT-29/OXA细胞对奥沙利铂的耐药性，其机制可能与细胞内LRP和P-gp表达降低有关。 |
| 关键词: 结肠癌 HT-29 奥沙利铂 多药耐药性 苦参碱 |
| DOI：10.3969/j.issn.1674-070X.2016.11.006 |
| Received:March 26, 2016 |
| 基金项目:上海市进一步加快中医药事业发展三年行动计划资助（2014-2016年，编号ZY3-CCCX-1-1008）。 |
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| The Reversal Effects of Matrine on the Multi-drug Resistance of Oxaliplatin in Human Colon HT-29 Cells and its Mechanism Research |
| ZHANG Dongmei,ZHANG Yaming |
| (Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200000, China) |
| Abstract: |
| Objective To study of matrine on the reversal of multi-drug resistance in HT-29/OXA cells and explore its underlying mechanisms. Methods Oxaliplatin resistant HT-29/OXA cells were established by gradually increasing the concentration of medicines. The reverse efficacy of Matrine on HT-29/OXA cells was determined by CCK-8 assay. The apoptosis and cell cycle were detected by flow cytometry. The expression levels of lung resistance protein (LRP) and P-glycoprotein (P-gp) mRNA and proteins was determined by quantitative real time PCR (qRT-PCR) and Western blot, respectively. Results Matrine can increase the sensibility of HT-29/OXA to oxaliplatin and reverse the part resisitance. Matrine (0.5 μg/mL) and matrine(3.0 μg/mL) alone had no effect on the cell cycle and apoptosis in HT-29/OXA cells. The oxaliplatinin combination with matrine significantly inhibited cell proliferation and induced apoptosis. Oxaliplatin(0.5 μg/mL) and matrine(3.0 μg/mL) alone had no effect on the expression level of LRP and P-gp mRNA and protein in HT-29/OXA cells. The expression of LRP and P-gp mRNA and protein was lower with drug combination. Conclusion Matrine could partially reverse the multi-drug resistance of HT-29/OXA cells to oxaliplatin, which may be related to the lower expression levels of LRP and P-gp. |
| Key words: colon cancer HT-29 oxaliplatin multi-drug resistance matrine |
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