Cullin蛋白4B在非小细胞肺癌中的表达及其临床意义和细胞功能分析

目的 探讨Cullin蛋白4B（CUL4B）在非小细胞肺癌（NSCLC）组织中的表达及其对NSCLC细胞生物功能的影响。方法 采用免疫组化、实时荧光定量PCR（qPCR）检测77例NSCLC组织及42例癌旁正常组织中的CUL4B表达。qPCR检测常见NSCLC细胞株（H358、H460、H1299、A549和SK-MES-1）中的CUL4B mRNA水平，选取表达最高的细胞株分别转染CUL4B siRNA（CUL4B干扰组）和CUL4B阴性对照siRNA（阴性对照组）的慢病毒载体，qPCR验证转染效率；采用MTT法检测干扰CUL4B表达24、48、72、96 h对细胞增殖的影响，流式细胞术和Transwell法检测干扰CUL4B表达48 h对细胞凋亡及侵袭能力的影响。结果CUL4B蛋白在NSCLC组织及癌旁正常组织中的高表达率分别为63.6%（49/77）和38.1%（16/42），差异有统计学意义（P<0.05）；CUL4B表达与肿瘤直径、淋巴结转移及临床分期均有关（P<0.05），而与性别、年龄、病理分型及组织分级无关（P>0.05）；qPCR检测NSCLC组织中CUL4B mRNA相对水平为2.713±0.246，经干扰CUL4B表达，NSCLC细胞增殖能力明显减弱，细胞凋亡率增加且侵袭转移能力降低。结论NSCLC组织及细胞株中CUL4B蛋白表达均明显升高，NSCLC组织CUL4B表达与淋巴结转移、临床分期及细胞分化有关；沉默CUL4B表达可影响NSCLC细胞的增殖、凋亡与侵袭。

Expression of Cullin 4B in non-small cell lung cancer and its clinical significance and cell function analysis

Objective To explore the expression of Cullin 4B （CUL4B） in non-small cell lung cancer （NSCLC） and its effect on NSCLC cell function. Methods The expression of CUL4B was detected by immunohistochemistry and quantitative real time polymerase chain reaction （qPCR） in 77 cases of NSCLC tissues and 42 cases of normal tissues adjacent to cancer. The qPCR was used to detect the mRNA level of CUL4B in common NSCLC cell lines， including H358， H460， H1299， A549 and SK-MES-1. The highest expression of the cell lines were transfected with siRNA CUL4B （CUL4B interference group） and CUL4B negative control siRNA （negative control group）， respectively. Transfection efficiency was verified by qPCR. The effect of interfering CUL4B expression on the cell proliferation at 24， 48， 72， 96 h was detected by MTT method. Flow cytometry and Transwell assay were used to detect the effect of CUL4B expression on cell apoptosis and invasion at 48 h. Results The high expression rates of CUL4B protein in NSCLC tissues and adjacent normal tissues were 636% （49/77） and 381% （16/42）， and the difference was statistically significant （P<0.05）. CUL4B expression was related to tumor diameter， lymph node metastasis and clinical stage （P<0.05）， but not related to gender， age， pathological type and histological grade （P>0.05）. The relative level of mRNA CUL4B in NSCLC tissue was 2.713±0.246， and interfering CUL4B expression of NSCLC cell， proliferation ability was significantly decreased， cell apoptosis increased and the invasion and metastasis ability decreased. Conclusion CUL4B protein expression was significantly increased in NSCLC tissues and cell lines， and was related to lymph node metastasis， clinical stage and cell differentiation. Silencing CUL4B can affect the proliferation， apoptosis and metastasis of NSCLC cells.