MTT比色法检测苦瓜MAP30蛋白对5637细胞增殖、MALAT1、HOTAIR及UCA1的影响  

Effect of MAP30 protein of Momordica charantia on proliferation,malat1,hotair and UCA1 of 5637 cells by MTT

作　　者：杨海霞[1] 张鹏[1] 陆伦[1] 邓秀英[1] 陈艳梅[1] YANG Haixia;ZHANG Peng;LU Lun;DENG Xiuying;CHEN Yanmei(Department of Pharmacy, Suining Central Hospital, Suining 629000, Sichuan, China)

机构地区：[1]遂宁市中心医院药学部,四川遂宁629000

出　　处：《西部医学》2021年第12期1862-1866,共5页Medical Journal of West China

摘　　要：目的探讨噻唑蓝(MTT)比色法检测苦瓜MAP30蛋白对5637细胞增殖、肺腺癌转移相关转录因子1(MALAT1)、长链非编码RNA HOX转录反义RNA(HOTAIR)基因及尿路上皮癌相关1(UCA1)的影响。方法按MTT法的相关操作原则与相关流程提取膀胱癌患者的苦瓜MAP30蛋白后进行克隆、表达及纯化处理,接着提取其处理膀胱癌5637细胞并进行MTT检测。探析苦瓜MAP30蛋白对5637细胞增殖和对MALAT1、HOTAIR及UCA1表达的相关性。结果抑制率:膀胱癌5637细胞增殖时所产生的抑制率越高时,MAP30蛋白浓度也越高,且时间越长,即MAP30蛋白浓度与膀胱癌5637细胞增殖呈正比(P<0.05)。凋亡率:膀胱癌5637细胞凋亡随MAP30浓度递增而逐渐上升,且MAP30浓度(培养48 h)为400μg/mL时,膀胱癌5637细胞凋亡率达到最高,MAP30浓度与膀胱癌5637细胞凋亡呈正比(P<0.05)。膀胱癌5637细胞lncRNA MALAT-1、HOTAIR、UCA1表达水平随苦瓜MAP30蛋白浓度的递增而逐渐降低,检测时间为48 h且苦瓜MAP30蛋白浓度为400μg/mL时上述三项指标的表达水平均显著低于48、24 h的其他苦瓜MAP30蛋白浓度(P<0.05)。结论苦瓜MAP30蛋白对膀胱癌5637细胞增殖的抑制和凋亡诱导方面有较好的效果,且可下调MALAT-1、HOTAIR、UCA1表达水平。Objective To investigate the effect of MAP30 protein of Momordica charantia on the proliferation,metastasis related transcription factor-1(MALAT1),long-chain noncoding RNA Hox transcriptional antisense RNA(HOTAIR)and urothelial carcinoma related 1(UCA1)of 5637 cells.Methods According to the relevant operating principles of MTT method,the MAP30 protein of balsam gourd(bladder cancer patients)was cloned,expressed and purified.Then 5637 bladder cancer cells were extracted and(MTT)was detected to explore the correlation between the proliferation of 5637 cells and the expression of MALAT1,HOTAIR and UCA1(real-time fluorescence quantitative polymerase chain reaction:RT-PCR).Results The higher the inhibition rate of bladder cancer 5637 cell proliferation,the higher the concentration of MAP30 protein.The longer the time,the longer the MAP30 protein concentration was proportional to the proliferation of bladder cancer 5637 cells(P<0.05).Apoptosis of bladder cancer 5637 cells increased gradually with increasing MAP30 concentration,and MAP30 concentration(48h)was 400μg/mL.The apoptosis rate of bladder cancer 5637 cells reached the highest level,and the MAP30 concentration was directly proportional to the proliferation of bladder cancer 5637 cells(P<0.05).The expression level of lncRNA MALAT 1,HOTAIR and UCA1 in bladder cancer 5637 cells gradually decreased with the increasing concentration of Momordica charantia MAP30 protein.The detection time was 48h and the concentration of bitter melon MAP30 protein was 400μg/mL.The expression levels of the above three indexes were significantly lower than that of other Momordica charantia MAP30 protein concentrations at 48h and 24h(P<0.05).Conclusion Balsam pear MAP30 protein has a good effect on inhibiting the proliferation of bladder cancer 5637 cells and inducing apoptosis,and can down-regulate the expression levels of MALAT-1,HOTAIR and UCA1.

关 键 词：MTT比色法 MAP30蛋白 5637细胞增殖 肺腺癌转移相关转录因子1 长链非编码RNA HOX转录反义RNA 尿路上皮癌相关1 

分 类 号：R737.14[医药卫生—肿瘤]