靶向MELK抑制食管鳞癌增殖与侵袭能力机制探讨  

Mechanism of inhibition of proliferation and invasion of esophageal squamous cell carcinoma by targeting MELK

作　　者：雷翔慧 林泽龙 陈丽青 胡玉林[1] LEI Xiang-hui;LIN Ze-long;CHEN Li-qing;HU Yu-lin(Department of Pathology,First People's Hospital of Chenzhou,Chenzhou 431000,China;Department of Oncology,Jinshazhou Hospital Affiliated to Guangzhou University of Traditional Chinese Medicine,Guangzhou 510400,China;Department of Oncology,Integrated Traditional Chinese and Western Medicine Hospital of Southern Medical University,Guangzhou 510400,China)

机构地区：[1]郴州市第一人民医院病理科,湖南郴州431000 [2]广州中医药大学附属金沙洲医院肿瘤科,广东广州510400 [3]南方医科大学中西医结合医院肿瘤科,广东广州510400

出　　处：《社区医学杂志》2022年第13期732-737,共6页Journal Of Community Medicine

基　　金：湖南省郴州市湘南学院课题(2019XJ81)

摘　　要：目的探讨母体胚胎亮氨酸拉链激酶(MELK)在食管鳞癌组织中的表达及其对食管鳞癌细胞增殖与侵袭能力的机制。方法免疫组化及蛋白质印迹法检测食管鳞癌癌组织与癌旁组织中MELK的表达;对食管鳞癌细胞株ECA109和KYSE510采用划痕实验和CCK8增殖实验,研究敲低MELK基因后食管鳞癌细胞株的侵袭能力、增殖能力及加入MELK靶向抑制剂OTS167后食管鳞癌细胞株的增殖能力。结果免疫组化和蛋白质印迹检测结果显示,MELK在食管鳞癌组织中高表达,而在癌旁组织或正常食管鳞状上皮组织不表达;划痕实验结果显示,在食管鳞癌细胞株ECA109和KYSE510中敲低MELK基因12 h后,食管鳞癌细胞ECA109和KYSE510实验组肿瘤细胞划痕愈合率均下降,但差异无统计学意义。敲低MELK基因36 h后,食管鳞癌细胞ECA109实验组肿瘤细胞划痕愈合率为(6.42±1.21)%,比对照组的(10.05±2.35)%明显下降,差异有统计学意义,t=4.540,P=0.038;食管鳞癌细胞KYSE510实验组肿瘤细胞划痕愈合率为(6.59±0.67)%,比对照组的(9.00±1.64)%明显下降,差异有统计学意义,t=4.720,P=0.006。CCK8细胞增殖实验结果显示,敲低MELK后,第4天食管鳞癌细胞ECA109实验组肿瘤细胞增殖能力D(450 nm)值为2.07±0.02,比对照组的2.57±0.03明显降低,差异有统计学意义,t=31.120,P=0.001;KYSE510实验组肿瘤细胞增殖能力D(450 nm)值为2.13±0.11,比对照组的2.38±0.02明显降低,差异有统计学意义,t=3.577,P=0.020。在食管鳞癌细胞株ECA109和KYSE510中加入MELK靶向抑制OTSSP167250μmol/L第5天后,食管鳞癌细胞ECA109实验组肿瘤细胞增殖能力D(450 nm)值为0.68±0.01,比对照组的2.67±0.14明显降低,差异有统计学意义,t=36.120,P=0.001;食管鳞癌细胞KYSE510实验组肿瘤细胞增殖能力D(450 nm)值为0.33±0.01,比对照组的2.78±0.09明显降低,差异有统计学意义,t=45.780,P=0.001。结论MELK在食管鳞癌中高表达,且起到促进肿瘤细胞的侵袭及增殖�Objective To investigate the expression of maternal and embryonic leucine zipper kinase(MELK)in esophageal squamous cell carcinoma and its mechanism of proliferation and invasion of esophageal squamous cells.Methods The expression of MELK in esophageal squamous cell carcinoma and adjacent tissues was detected by immunohistochemistry and Western blot.The invasiveness and proliferation of esophageal squamous cell carcinoma cell lines ECA109 and KYSE510 after MELK gene knockdown and the proliferation of esophageal squamous cell carcinoma cell lines after MELK targeted inhibitor OTS167 were studied by scratch test and CCK8 proliferation test.Results Immunohistochemistry and Western blot showed that MELK was highly expressed in esophageal squamous cell carcinoma,but not in adjacent tissues or normal esophageal squamous epithelium.The results of scratch test showed that after knocking down MELK gene in esophageal squamous cell carcinoma cell lines ECA109 and KYSE510 for 12 hours,the scratch healing rate of tumor cells in the experimental groups of esophageal squamous cell carcinoma cells ECA109 and KYSE510 decreased,but the difference was not statistically significant.After 36hours,the scratch healing rate of esophageal squamous cell carcinoma cell ECA109in the experimental group was(6.42±1.21)%,which was significantly lower than that in the control group(10.05±2.35)%,and the difference was statistically significant,t=4.540,P=0.038.The scratch healing rate of tumor cells in the experimental group of esophageal squamous cell carcinoma KYSE510was(6.59±0.67)%,which was significantly lower than that in the control group(9.00±1.64)%,with a statistically significant difference,t=4.720,P=0.006.The results of CCK8cell proliferation experiment showed that after MELK knockdown,the proliferative capacity D(450nm)of esophageal squamous cell carcinoma cell ECA109in the experimental group was 2.07±0.02on the 4th day,which was significantly lower than that in the control group(2.57±0.03),the difference was statistically signifi

关 键 词：母体胚胎亮氨酸拉链激酶 食管鳞癌 靶向 侵袭 增殖 

分 类 号：R735.1[医药卫生—肿瘤]