摘要
目的 探讨96序列相似的家庭成员B（FAM96B）在肝细胞肝癌中的表达,以及对人肝癌细胞株HepG2增殖和凋亡的影响。方法 分别采用实时荧光定量聚合酶链反应（FQ-PCR）及Western blot技术检测FAM96B在42例肝癌肿瘤组织和癌旁组织、HepG2和L-02细胞中的mRNA及蛋白表达量,比较其差异。构建pcDNA3.1-myc-FAM96B真核表达载体,并通过脂质体质粒转染HepG2细胞,采用噻唑蓝（MTT）法及流式细胞仪（FACS）检测FAM96B基因对HepG2增殖及凋亡的影响。结果 42例肝癌组织中,Real-time PCR结果显示FAM96B在肿瘤组织中的mRNA表达量是癌旁组织的（0.304±0.094）倍,且其在HepG2细胞中的mRNA表达量是L-02细胞的（0.406±0.115）倍,差异有统计学意义（P〈0.05）。Western blot结果显示FAM96B在肿瘤组织中的蛋白表达是癌旁组织的（0.288±0.132）倍（P〈0.05）,且其在HepG2细胞中的蛋白表达是L-02细胞的（0.359±0.143）倍（P〈0.05）。MTT细胞实验结果显示FAM96B组吸光度值显著低于对照组（P〈0.05）,即过表达FAM96B可抑制HepG2细胞增殖。流式检测显示过表达FAM96B组的HepG2细胞凋亡率（33.3%）与对照组（3.3%）比较增高,差异有统计学意义（P〈0.05）。结论 FAM96B在42例肝癌肿瘤组织中的表达显著低于癌旁组织,并且其在HepG2细胞中的表达低于L-02细胞,过表达FAM96B可抑制HepG2细胞增殖并诱导其凋亡。该基因可能在肝细胞肝癌的发生发展中发挥重要作用。
Objective To study the expression difference of family with sequence similarity 96, member B （FAM96B） in human hepatocellular carcinoma （HCC） and its impact on the proliferation and apoptosis of HepG2 cell line. Methods FAM96B mRNA and protein expression was examined respectively by real-time fluorogenic quantitative polymerase chain reacton （FQ-PCR） and Western blotting in tumor tissues and adjacent tissues from 42 patients with HCC. The FAM96B mRNA and protein expression was detected in HepG2 and L-02 cells. Moreover the eukaryotic vector of human FAM96B was constructed and transfected into HepG2 cells. The proliferative rate and apoptosis rate of HepG2 cells were examined by the methyl thiazol tetrazolium （MqT） assay and fluorescence-activated cell sorting （FACS）. Results In 42 patients with HCC, the global mRNA expression of FAM96B was 0. 304 ± 0. 094 times in tumor tissues than in adjacent tissues . And its mRNA expression was 0. 406 ±0. 115 times in HepG2 cell than in L-02 cell. While the Western blotting analysis confirmed that FAM96B protein expression in tumor tissues was O. 288 ± O. 132 times lower than in the adjacent tissues （P 〈 0. 05 ）. And FAM96B protein expression in HepG2 cell was 0. 359 ± 0. 143 times lower than in L-02 cell. The Mq＇T assay showed that the A value in FAM96B group was lower than in control group, and overexpression of FAM96B significantly inhibited proliferation of HepG2 cells （ P 〈 0. 05 ）. FACS analysis indicated that there was significant difference in cell apoptosis rate between FAM96B-expressing clones group and the control group （vector-transfected cells group） （P 〈 0. 05 ）. Conclusion FAM96B expression showed statistically significant difference between tumor tissues and adjacent tissues in 42 cases of HCC. Moreover FAM96B expression in HepG2 cells was significantly lower than in L-02 cells （P 〈 0. 05）. Furthermore, overexpression of FAM96B can induce ap- optosis of HepG2 cells and inhibit proliferation of HepG2 cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第7期1489-1491,共3页
Chinese Journal of Experimental Surgery
基金
湖北省自然科学基金资助项目（2012FFB06805）
关键词
癌
肝细胞
96序列相似的家庭成员B基因
增殖
Carcinoma, hepatocellular
Family with sequence similarity 96, member B
Pro-liferation
