CAJ | 학술논문

目的：肿瘤耐药的产生是肿瘤治疗失败的主要原因之一,α烯醇化酶( eno1)与肿瘤细胞耐药产生和发展密切相关。探究eno1对人慢性粒细胞白血病耐药细胞株 K562/A02生长及耐药的影响。方法筛选3株稳定干扰eno1的细胞系K562/A02-sheno1和对照细胞系K562/A02-shcon；采用细胞计数法测定细胞生长速率, MTT法测定细胞增殖能力,细胞内罗丹明123含量测定细胞外排药物能力,real-time PCR反应测定基因mRNA表达水平, Western blot实验测定基因蛋白表达水平。结果与敏感性细胞株 K562相比, eno1在耐药细胞株K562/A02中为高表达状态,其在mRNA水平和蛋白水平表达分别增高(2．85±0．56)倍和(1．43±0．05)倍；而K562/A02细胞生长速率与K562细胞相比差异无显著性。 K562/A02-sheno1细胞系与对照组 K562/A02-shcon相比生长速率降低,对抗肿瘤药物紫杉醇和阿霉素的敏感性均增强,且K562/A02-sheno1细胞系中罗丹明123含量也明显增高。 K562/A02-sheno1细胞系中耐药相关基因MDR1表达水平降低。结论稳定干扰 eno1表达能抑制K562/A02细胞生长,有效逆转K562/A02细胞耐药性,提高其对药物的敏感性,其机制与MDR1基因表达相关。
목적：종류내약적산생시종류치료실패적주요원인지일,α희순화매( eno1)여종류세포내약산생화발전밀절상관。탐구eno1대인만성립세포백혈병내약세포주 K562/A02생장급내약적영향。방법사선3주은정간우eno1적세포계K562/A02-sheno1화대조세포계K562/A02-shcon；채용세포계수법측정세포생장속솔, MTT법측정세포증식능력,세포내라단명123함량측정세포외배약물능력,real-time PCR반응측정기인mRNA표체수평, Western blot실험측정기인단백표체수평。결과여민감성세포주 K562상비, eno1재내약세포주K562/A02중위고표체상태,기재mRNA수평화단백수평표체분별증고(2．85±0．56)배화(1．43±0．05)배；이K562/A02세포생장속솔여K562세포상비차이무현저성。 K562/A02-sheno1세포계여대조조 K562/A02-shcon상비생장속솔강저,대항종류약물자삼순화아매소적민감성균증강,차K562/A02-sheno1세포계중라단명123함량야명현증고。 K562/A02-sheno1세포계중내약상관기인MDR1표체수평강저。결론은정간우 eno1표체능억제K562/A02세포생장,유효역전K562/A02세포내약성,제고기대약물적민감성,기궤제여MDR1기인표체상관。
Aim Drug resistance is one of the major hinders on cancer treatments. α-enolase ( eno1 ) was closely related to the generation and development of drug resistance. This article aims to study the effect of eno1 on cell growth and drug resistance in human chro-nic myeloid leukemia cell line K562/A02 . Methods We screened three eno1 stable silencing cells K562/A02-sheno1 and its control cells K562/A02-shcon. Cell count assay was performed to test cell growth, MTT assay was used to test cell proliferation, flow cytometry was used to test the intra-cellular Rho123 content, the expression of genes were tested by real-time PCR assay and western blot assay on mRNA level and protein level, respectively. Results eno1 was o-ver-expressed in K562/A02 cells and its expression was increased by ( 2. 85 ± 0. 56 ) times and ( 1. 43 ± 0. 05 ) times on mRNA level and protein level com-pared to K562 cells. However, there was no difference in cell growth rate between K562/A02 cells and K562 cells. K562/A02-sheno1 cells showed lower cell growth rate and higher drug sensitivity to anti-cancer drugs taxol and doxorubicin. Moreover the Rho123 content was increased in K562/A02-sheno1 cells. The expression of MDR1 decreased in both mRNA level and protein level in K562/A02-sheno1 cells. Conclusion eno1 silencing could suppress cell growth, reverse drug resistance and increase its drug sensitivity in K562/A02 cells, and the mechanism was associated with the MDR1 gene.