大黄素通过下调多药耐药基因-1的表达改善胰腺癌细胞对吉西他滨的耐药研究

Effect of emodin on the improvement of drug resistance of gemcitabine in pancreatic cancer cell line by down-regulating the expression of multidrug resistance gene-1

目的探讨大黄素改善胰腺癌细胞株SW1990/Gemcitabine(GEM)耐药作用及其可能机制。方法用吉西他滨体外浓度递增法诱导培养人胰腺癌细胞株10个月,获得耐药细胞株SW1990/GEM。将细胞分为4组:对照组、大黄素组、吉西他滨组、联合组。对照组,加入等量0. 1%DMSO溶液;其他3组分别给予10μmol·L^(-1)大黄素、20μmol·L^(-1)吉西他滨与联合两药(大黄素组+吉西他滨)来分别处理SW1990/GEM细胞。用MTT法检测细胞增殖,以流式细胞仪检测细胞凋亡,以逆转录-聚合酶链反应实验检测细胞中MDR^(-1)基因的表达水平,以流式细胞技术检测P-gp的功能。罗丹明外排实验检测SW 1990/GEM细胞内P-gp的功能。结果联合组与吉西他滨相比,可明显抑制胰腺癌SW1990/Gemcitabine细胞的增殖。吉西他滨单独处理SW 1990/GEM与SW1990细胞时,对2个细胞的抑制率分别为13. 34%与36. 52%,两者比较差异有统计学意义(P <0. 05),说明与细胞株SW 1990相比较,SW 1990/GEM细胞株对吉西他滨具备了明显的耐药性。当吉西他滨联合大黄素处理细胞时,对2种细胞的抑制率分别是40. 45%与43. 87%,差异不明显,由此可见大黄素能够增强吉西他滨对耐药细胞株SW 19901GEM增殖的抑制作用。联合组与吉西他滨组比较,可抑制细胞MDR^(-1)基因的表达,差异有统计学意义(P <0. 05)。结论大黄素通过下调多药耐药基因^(-1)的表达,可以改善胰腺癌细胞对吉西他滨的耐药。

Objective To investigate the effect of emodin on the gemcitabine- resistant pancreatic cancer cell line SW1990/ Gemcitabine （GEM）, and explore the potential mechanism of its action. Methods The pancreatic cancer cell line SW1990 was treated by intermittently increasing the concentration of gemcitabine in the culture medium for 10 months, and SW1990/GEM cells were obtained. This experiment was divided into control group, emodin group, gemcitabine group and combination group （emodin ＋ gemcitabine）. SW1990/GEM and SW1990 cells were treated with emodin （ 10 μmol· L^-1 ） and gemcitabine （20 μmol · L^- 1 ） alone or those two together in three groups for 48 h, in con- trol group cells were treated with 0. 1% DMSO for 48 h. Cell proliferation was analyzed by MTr. Reverse transcription- polymerase chain reaction was performed to analyze the protein and gene expression of muhidrug resistance gene - 1 （ MDR - 1 ）. Flow cytometric was applied to analyze the function of the P - glycoprotein （ P - gp）. The Rhodamine 123 efflux experiment was applied to assay P - gp function in SW1990/Gemcitabine cells. Results Compared with gemcitabine group, the combination group could significantly inhibit the proliferation of SW1990/GEM ceils. Treatment of SW1990/GEM and SW1990 cells with gemcitabine alone could inhibit 13.34% and 36. 52 % of cell viability, there was significant difference between the two group （P 〈 0. 05 ） The results showed that the SW1990 / GEM cell line had an obvious resistance to gemcitabine compared with the cell line SW 1990. While SW 1990/GEM and SW 1990 ceils were treated with gemcitabine combined with emodin, cell viability was inhibited to 40. 45% and 43.87% , there was no could enhance the anti - proliferative effect of gemcitabine significant difference between the two group. The emodin on drugresistance cell SW1990/GEM. Compared with gemcitabine group, the combination group could significantly inhibit the expression of MDR - 1 gene, and the difference was statistically significant （P 〈 0. 05 ）. Conclusion Effect of emodin can down - regulate the expression of MDR - 1 and then improve the drug resistance of gemcitabine in pancreatic cancer.