高压电场对A549细胞中ABCG2和V-ATPase的表达及其耐药性的影响

目的:探讨高压电场对A549细胞中ABCG2和V-ATPase表达量的影响;探讨高压电场对A549细胞耐药性的影响。方法:MTT法测细胞生长曲线,明确能导致细胞可逆性电穿孔的最高电场强度。慢病毒构建ABCG2和V-ATPase低表达的A549细胞系,并用电场处理,用q-RT-PCR和Western-blot法检测处理前后ABCG2和V-ATPase的m RNA和蛋白表达量的变化。最适强度的高压电场处理各组细胞,在处理前后的细胞中分别加入阿霉素,用高效液相色谱法检测各组细胞中阿霉素浓度。结果:当电场强度为1500 V/cm时,肿瘤细胞增殖最慢;电场强度为1500 V/cm时,肿瘤细胞中ABCG2和V-ATPase的m RNA和蛋白的表达量分别降至对照组的58%和61%,具有统计学差异;1500 V/cm强度的电场可以提高肿瘤细胞内阿霉素的浓度3-4倍。结论:高压电场可以显著降低肿瘤细胞中V-ATPase和ABCG2的m RNA和蛋白的表达量并降低肿瘤细胞的耐药性。

Effects of High Voltage Electric Field on the Expression of ABCG2 and V-ATPase in A549 Cells and Drug Resistance of A549 Cells

ABSTRACT Objective: To investigate the effect of high voltage electric field on the expression of ABCG2 and V-ATPase in A549 cells, and to explore the effect of high voltage electric field on drug resistance of A549 cells. Methods: The cell growth curves was made by using MTT method to determine the highest electric field strength which can lead to reversible cell electroporation. A549 cell line was treated with virus to downregulation the expression of ABCG2 and V-ATPase. Then, treating each groups with high voltage electric field, the changes of the expression of mRNA and protein of ABCG2 and V-ATPase were detected by using q-RT-PCR and Western-blot methods before and after treatment. Cells were treated with the perfect strength high voltage electric field, adriamycin were added in cells before and after treatment, high performance liquid chromatography method was used to detect adriamycin concentration in each groups. Results: When the electric field intensity is 1500V/cm, the proliferation of tumor cells is the slowest and the expression levels of ABCG2 and V-ATPase mRNA in tumor cells decreased to 58% and 61% of the control group, there are statistically significant differences between groups. Then, electric field strength of 1500V/cm can significantly improve the concentration of adriamycin in tumor cells by three to four times. Conclusion: High voltage electric field can significantly reduce the expression of mRNA and protein of V-ATPase and ABCG2 in tumor cells and reduce the drug resistance of tumor cells.