摘要
目的分析肝细胞癌（HCC）中雄激素受体（AR）、三磷酸腺苷酶家族蛋白2（ATAD2）的表达与患者临床指标的关系，探讨二氢睾酮（DHT）、AR与ATAD2对肝癌细胞增殖的影响。方法收集2012年2月至12月中国医科大学附属第一医院75例HCC患者标本及临床资料。LM3和Huh7细胞随机分为对照组、DHT组、DHT＋CDX（比卡鲁胺）组、CDX组；LM3和Huh7细胞均分为Ri-ATAD2组（加入干扰片段）和Ri-C组（加入对照载体序列）。免疫组化检测AR、ATAD2表达，并分析其与临床指标及患者生存的关系。Real-time PCR、Western印迹检测AR、ATAD2表达；CCK-8检测各组细胞增殖。结果HCC患者按AR和ATAD2表达高低分组。与31例AR低表达比较，44例高表达患者肿瘤≤5 cm比例升高，TNM分期Ⅰ＋Ⅱ比例降低；与35例ATAD2低表达比较，40例高表达患者转移和肿瘤分化Ⅲ＋Ⅳ级比例升高，差异有统计学意义（P〈0.05）。ATAD2高表达患者术后总生存率低于其他患者，差异有统计学意义（P〈0.05）。多因素Cox回归分析显示ATAD2表达（HR＝1.935，95%CI：1.066～3.515）、转移（HR＝2.212，95%CI：1.059～4.619）为患者术后预后不良独立预测指标。与正常人肝细胞LO2比较，LM3及Huh7组AR、ATAD2蛋白和mRNA相对表达升高，差异有统计学意义（P〈0.05）。加入DHT后，与对照组比较，肝癌细胞在48、72小时后增殖率明显升高，差异有统计学意义（P〈0.05）。加入CDX后，DHT诱导的LM3及Huh7增殖受到抑制。DHT可诱导ATAD2的蛋白表达，而CDX可抑制ATAD2的表达。肝癌细胞加入DHT 48、72小时后，与Ri-C组比较，Ri-ATAD2组增殖下降，差异有统计学意义（P〈0.05）。结论DHT及AR可通过诱导ATAD2的表达进一步促进肝癌细胞的增殖。ATAD2可能是DHT及AR促进肝癌细胞增殖的潜在机制之一。
Objective To investigate the expression of androgen receptor（AR）, ATAD2 in hepatocellular carcinoma（HCC） and the correlations with clinicopathological features, and the role of DHT/AR and ATAD2 in proliferation of HCC cells.Methods The samples of 75 patients with HCC in the First Affiliated Hospital of China Medical University from February 2012 to December 2012 were collected. LM3 and Huh7 cells were divided into control group, DHT group, DHT ＋ CDX （bicalutamide） group and CDX group; and also divided into Ri-ATAD2 group （adding interference fragments） and Ri-C group （adding control vector sequence）. Immunohistochemistry was used to detect the expression of AR and ATAD2, and to analyze the correlations between clinical features and survival of patients. Real-time PCR and Western Blot were used to detect the expression of AR and ATAD2, and CCK-8 was used to detect cell proliferation.Results HCC patient samples were grouped according to AR and ATAD2 expression. Compared with low AR expression group （n=31）, the ratio of tumor 〈5 cm in high expression group （n=44） was higher, and the ratio of TNM stage I ＋ II was lower. Compared with low ATAD2 expression group （n=35）, the ratio of metastasis and tumor differentiation grade III ＋ IV was higher in high expression group （n=40）, and the difference was statistically significant （P〈0.05）. The overall survival rate of patients with high expression of ATAD2 was lower than other patients, and the differences were statistically significant （P〈0.05）. Multivariate Cox regression analysis showed that ATAD2 expression （HR=1.935, 95% CI： 1.066-3.515） and metastasis （HR=2.212, 95% CI： 1.059-4.619） were independent predictors of poor prognosis. Compared with LO2 cells, the mRNA and protein level of AR and ATAD2 in LM3 and Huh7 cells were significantly higher, and the differences were statistically significant （P〈0.05）. And the proliferation rate of HCC cells increased significantly after 48 and 72 hours compare
作者
王开宇
李文心
马作红
尚海
郝志强
冯照强
华向东
Wang Kaiyu;Li Wenxin;Ma Zuohong;Shang Hai;Hao Zhiqiang;Feng Zhaoqiang;Hua Xiangdong(Department of Hepatobiliary Pancreatic Surgery,Cancer Hospital of China Medical University,Liaoning Cancer Hospital,Shenyang 110042,China)
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2018年第11期742-746,共5页
Chinese Journal of Hepatobiliary Surgery
关键词
二氢睾酮
雄激素受体
三磷酸腺苷酶家族蛋白2
肝细胞癌
增殖
Dihydrotestosterone
Androgen recceptor
ATPase family AAA domain-containing protein 2
Hepatocellular carcinoma
Proliferation
