摘要
目的探讨二氢青蒿素(DHA)联合顺铂(CDDP)对人胃癌SGC7901细胞增殖和耐药因子表达的影响。方法将体外培养的人胃癌SGC7901细胞株和人胃黏膜上皮细胞GES-1细胞株分为6组,即SGC7901对照组(0.1%二甲基亚砜100μl)、SGC7901 DHA组(25μmol/L DHA 100μl)、SGC7901 CDDP组(2.5μg/ml CDDP 100μl)、SGC7901 DHA+CDDP组(25μmol/L DHA和2.5μg/ml CDDP各100μl)、GES-1对照组(0.1%二甲基亚砜100μl)和GES-1 DHA组(25μmol/L DHA 100μl)。采用磺酰罗丹明染色法检测各组细胞增殖的抑制情况,以光密度(OD)值表示;反转录聚合酶链反应检测耐药因子多药耐药基因1(MDR1)、多药耐药相关蛋白1(MRP1)、拓扑异构酶Ⅱ(TopoⅡ)、谷胱甘肽转移酶-π(GST-π)、B细胞淋巴瘤/白血病蛋白-2(Bcl-2)、Bcl-2相关X蛋白(Bax)mRNA水平,以各目的基因片段密度值/内参照β-actin密度值表示;蛋白质印迹法检测上述耐药因子蛋白水平,以各目的蛋白条带吸光度值/内参照β-actin吸光度值表示。结果 SGC7901对照组、SGC7901 DHA组、SGC7901CDDP组、SGC7901 DHA+CDDP组OD值分别为(0.77±0.14)、(0.58±0.13)、(0.52±0.12)、(0.30±0.05),差异有统计学意义(F=18.71,P<0.05);SGC7901 DHA组、SGC7901 CDDP组及SGC7901 DHA+CDDP组OD值与SGC7901对照组比较,差异亦有统计学意义(t值分别为3.60、4.05和6.81,P<0.05)。GES-1对照组OD值为(0.79±0.13),GES-1 DHA组为(0.78±0.12),差异无统计学意义(t=-1.24,P>0.05)。SGC7901对照组、SGC7901 DHA组、SGC7901 CDDP组、SGC7901 DHA+CDDP组各耐药因子mRNA和蛋白水平比较,差异均有统计学意义(P<0.05);SGC7901 DHA组、SGC7901 CDDP组及SGC7901 DHA+CDDP组MDR1、MRP1、GST-π、Bcl-2mRNA和蛋白水平较SGC7901对照组降低,差异均有统计学意义(P<0.05);TopoⅡ和Bax mRNA和蛋白水平较SGC7901对照组升高,差异亦有统计学意义(P<0.05)。GES-1对照组与GES-1 DHA组各耐药因子mRNA和蛋白水平比较,差异均无统计学意义(P>0.05)。结论 DHA对胃癌SGC7901细胞生长具有明显的抑制作用,可增强化疗药物CDDP对SGC7901细胞增殖抑制作用,并能通过不同途径特异性抑制胃癌SGC7901细胞耐药因子的表达。
Objective To investigate the effect of dihydroartemisinin （DHA） combined with cisplatin （CDDP） on the proliferation and expression of muhidrug resistance associated factors in human gastric cancer SGC7901 cells. Methods Human gastric cancer SGC7901 cell strains and human gastric mucosa epithelial GES - 1 cell strains cultured in vitro were divided into six groups, namely SGC7901 control group （0. 1% DMSO 100μl）, SGC7901 DHA group （25 μmol/L DHA 100 p]）, SGC7901 CDDP group （2. 5μg/ml CDDP 100 p.1）, SGC7901 DHA ＋ CDDP group （25 μmol/L DHA and 2. 5 μg/ml CDDP with each 100 p]）, GES- 1 control group （0. 1% DMSO 100μl） and GES -1 DHA group （25μmol/L DHA 100 μl）. Sul- forhodamine staining was used to test cell proliferation of each group and the results were expressed in OD. RT - PCR was used to test mRNA of MDRI, MRP1, Topo Ⅱ , GST - π, Bcl - 2 and Bax and the results were expressed in specific fragment density/ - actin density. Western blot was used to test the protein level of the above mentioned muhidrug resistance associated factors and the results were expressed in specific protein bands optical density/β - actin optical density. Results The values of OD in SGC7901 control group, SGC7901 DHA group, SGC7901 CDDP group and SGC7901 DHA ＋ CDDP group were （0.77 ±0. 14）, （0.58 ±0. 13）, （0.52 ±0. 12） and （0.30±0.05） respectively, and the difference were statistically significant （F = 18.71, P 〈0. 05） . The OD values in SGC7901 DHA group, SGC7901 CDDP group and SGC7901 DHA ＋ CDDP group showed statistically significant differences compared with that of the SGC7901 control group （ t = 3.60, 4. 05 and 6. 81, P 〈 0. 05） . The OD values of GES - 1 control group and GES - 1 DHA group were （0. 79 ±0. 13） and （0. 78 ±0. 12） respectively, and the difference was not statistically significant （ t = - 1.24, P 〉 0. 05 ） . The mRNA and protein levels of each muhidrug resistance associated factor in SGC7901 control group, SGC7901 DHA group, SGC7901 CDDP group and SGC7901 DHA ＋ CDDP group showed statistically significant differences （ P 〈 0. 05 ） . The mRNA and protein levels of MDR1, MRP1, GST -π and Bcl -2 in SGC7901 DHA group, SGC7901 CDDP group and SGC7901 DHA ＋ CDDP group were significantly reduced compared with those of the SGC7901 control group （P 〈0. 05）, but the mRNA and protein levels of Tope ]1 and Bax were signifi- cantly elevated compared with those of the SGC7901 control group （P 〈 O. 05 ） . The mRNA and protein levels of each muhidrug resistance associated factor in GES - 1 control group and GES - 1 DHA group showed no statistically significant differences （ P 〉 0. 05）. Conclusion DHA can obviously inhibit the growth of gastric cancer SGC7901 cells. It can enhance the inhibition effect on SGC7901 ceils proliferation by CDDP and has specific inhibition effect on the expression of muhidmg resistance associated factors of SGC7901 in different ways.
出处
《中国全科医学》
CAS
CSCD
北大核心
2012年第27期3130-3134,共5页
Chinese General Practice
关键词
青蒿素类
顺铂
胃肿瘤
细胞增殖
抗药性
肿瘤
Artemisinins
Cisplatin
Stomach neoplasms
Cell proliferation
Drug resistance, neoplasms
作者简介
通讯作者：王爱军，063000河北省唐山市工人医院肿瘤外科；E—mail：waits@126．com
