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[小干扰RNA干扰β-连环蛋白表达对人多发性骨髓瘤细胞系多药耐药的影响]

[Effect of siRNA-Interfering β-Catenin Expression on MDR of Human Multiple Myeloma Cell Line].

作者信息

Zhang Jian-Guo, Li Yu-Peng, Fang Zi-Qing, Huang Xing-Ru, Chen Xu, Fang Jian

机构信息

Third Clinical Medical College of Guangdong University of Traditional Chinese Medicine, Guangzhou 510000, Guangdong Province, China.

Department of Orthopedics, The Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510000, Guangdong Province, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2019 Apr;27(2):477-481. doi: 10.19746/j.cnki.issn.1009-2137.2019.02.027.

Abstract

OBJECTIVE

To investigate the effecr of siRNA-interfering β-catenin expression on drug-resistance of multiple myeloma cells.

METHODS

The multiple myeloma cell line RPMI-8226 was cultured in vitro. The maphalan-resistant cell model was established by concentration gradient ascending of durg, then the drug-resistant cell line was instantaneously transfected with β-catenin siRNA, the sensitivity of RPMI 8226 cells to maphalan was detected by CCK-8 meltod before and after the transfection with siRNA; the mRNA and protein expression of β-catenin was detected by qRT-PCR and Western blot respectively, the apoptosis of cells was detected by flow cytometry.

RESULTS

IC of maphalan decreased from (5.29±0.19) μmol/L to (1.88±0.64) μmol/L, suggesting that the deplation of β-eatenin restored the sensitivity of drug-resistant cell line RPMI-8226 to malphalan. The Western blot showed that after the instaintaneous transfection with β-catenin siRNA, the β-catenin protein expression level obviously decreased, compared with level before transfection. After transfection, the maplalan-inducing apoptosis rate of cells increased from (35±0.5)% to (54±0.4)%, suggesting that the β-catinin gene may correlated with drug-resistance of cells. Interfering the expression of β-catenin gene could enhance the sensitivity of drug-resistant RPMI-8226 cells to maphalan.

CONCLUSION

The β-catenin siRNA interfereuce can inhisit the β-catenin gene expression in Wnt/β-catenin signaling pathway, suppress the cell proliferation, enhence the toxicity of maphalan on drug-resistant RPMI-8226 cells, thus result in increase of cell apoptosis.

摘要

目的

探讨小干扰RNA(siRNA)干扰β-连环蛋白表达对多发性骨髓瘤细胞耐药性的影响。

方法

体外培养多发性骨髓瘤细胞系RPMI-8226。通过药物浓度梯度递增建立马法兰耐药细胞模型,然后用β-连环蛋白siRNA瞬时转染耐药细胞系,用CCK-8法检测转染siRNA前后RPMI 8226细胞对马法兰的敏感性;分别用qRT-PCR和蛋白质免疫印迹法检测β-连环蛋白的mRNA和蛋白表达,用流式细胞术检测细胞凋亡情况。

结果

马法兰的半数抑制浓度(IC)从(5.29±0.19)μmol/L降至(1.88±0.64)μmol/L,提示β-连环蛋白缺失恢复了耐药细胞系RPMI-8226对马法兰的敏感性。蛋白质免疫印迹法显示,用β-连环蛋白siRNA瞬时转染后,β-连环蛋白蛋白表达水平明显低于转染前。转染后,马法兰诱导的细胞凋亡率从(35±0.5)%升至(54±0.4)%,提示β-连环蛋白基因可能与细胞耐药性相关。干扰β-连环蛋白基因表达可增强耐药RPMI-8226细胞对马法兰的敏感性。

结论

β-连环蛋白siRNA干扰可抑制Wnt/β-连环蛋白信号通路中β-连环蛋白基因表达,抑制细胞增殖,增强马法兰对耐药RPMI-8226细胞的毒性,从而导致细胞凋亡增加。

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