探讨HBx上调肝细胞癌（肝癌）中高表达的长链非编码RNA （lncRNA-HEIH）对肝癌细胞增殖的影响。

标本来源于2015年9月至2016年3月中山大学附属第三医院和中山大学附属肿瘤医院30例HBV相关性肝癌患者手术组织。所有患者均签署知情同意书，符合医学伦理学规定。其中男24例，女6例；年龄20～65岁，中位年龄52岁。采用RT-PCR检测30例患者肝癌组织及癌旁组织中HBx、lncRNA-HEIH mRNA的表达水平。构建稳定表达HBx的肝癌细胞系SMMC-7721-HBx及Bel-7404-HBx（实验细胞），及其对照细胞SMMC-7721-Vector和Bel-7404-Vector。RT-PCR及Western blot检测细胞株lncRNA-HEIH的表达。根据有否转染siRNA-HEIH，将实验细胞分别分为干扰组和非干扰组。MTT检测两组吸光度值（A₄₉₀）。实验数据比较采用t检验，lncRNA-HEIH与HBx表达相关性检验采用Pearson相关。

肝癌组织中HBx、lncRNA-HEIH mRNA表达量分别为0.91±0.04、0.93±0.05，明显高于癌旁组织的0.53±0.01、0.35±0.02（t=7.95，10.42；P<0.05）。lncRNA-HEIH mRNA与HBx mRNA的表达水平呈正相关（r=0.840，P<0.05）。实验细胞SMMC-7721-HBx和Bel-7404-HBx中lncRNA-HEIH表达量分别为1.05±0.22、1.20±0.17，明显高于对照细胞的0.45±0.10、0.50±0.12（t=5.69，3.78；P<0.05）。SMMC-7721-HBx和Bel-7404-HBx干扰组48、72、96 h的A₄₉₀分别为0.26±0.05、0.38±0.04、0.43±0.10和0.27±0.05、0.30±0.12、0.39±0.05，明显低于非干扰组的0.37±0.07、0.59±0.12、0.80±0.10和0.38±0.07、0.62±0.12、0.78±0.11 （t=-5.72，-3.47，-3.82和-3.89，-5.57，-8.34；P<0.05）。

lncRNA-HEIH高表达可能与HBV相关性肝癌发生有关，HBx基因可通过上调lncRNA-HEIH的表达参与HBx基因促进肝癌细胞增殖。

To investigate the effect of up-regulating long noncoding RNA high expression in hepatocellular carcinoma (lncRNA-HEIH) by HBx on the proliferation of hepatocellular carcinoma (HCC) cells.

The samples were collected from 30 patients with hepatitis B virus (HBV)-related HCC who received surgery in the Third Affiliated Hospital of Sun Yat-sen University and Cancer Center of Sun Yat-sen University between September 2015 and March 2016. The informed consents of all patients were obtained and the local ethical committee approval was received. Among 30 patients, 24 were males and 6 were females, aged 20-65 years old with a median age of 52 years old. The expression levels of HBx mRNA and lncRNA-HEIH mRNA in the HCC tissues and paracarcinoma tissues were detected by RT-PCR. HCC cell lines SMMC-7721-HBx and Bel-7404-HBx stably expressing HBx were established as the experimental cells. And the control cells SMMC-7721-Vector and Bel-7404-Vector were also established. The expression of lncRNA-HEIH in the cell lines was detected by RT-PCR and Western blot. According to siRNA-HEIH was transfected or not, the experimental cells were respectively divided into the interference group and the non-interference group. The absorbance values (A₄₉₀) of two groups were detected by MTT. The experimental data were compared using t test. The relationship between the expression of lncRNA-HEIH and HBx was analyzed using Pearson correlation analysis.

The expression levels of HBx mRNA and lncRNA-HEIH mRNA in the HCC tissues were respectively 0.91±0.04 and 0.93±0.05, significantly higher than 0.53±0.01 and 0.35±0.02 in the paracarcinoma tissues (t=7.95, 10.42; P<0.05). The expression level of lncRNA-HEIH mRNA was positively correlated with that of HBx mRNA (r=0.840, P<0.05). The expression level of lncRNA-HEIH in the SMMC-7721-HBx and Bel-7404-HBx cells was respectively 1.05±0.22 and 1.20±0.17, significantly higher than 0.45±0.10 and 0.50±0.12 in the control cells (t=5.69, 3.78; P<0.05). The A₄₉₀ in the SMMC-7721-HBx and Bel-7404-HBx interference groups was respectively 0.26±0.05, 0.38±0.04, 0.43±0.10 and 0.27±0.05, 0.30±0.12, 0.39±0.05 at 48, 72 and 96 h, significantly lower than 0.37±0.07, 0.59±0.12, 0.80±0.10 and 0.38±0.07, 0.62±0.12, 0.78±0.11 in the non-interference group (t=-5.72, -3.47, -3.82 and -3.89, -5.57, -8.34; P<0.05).

High expression of lncRNA-HEIH is probably correlated with the incidence of HBV-related HCC. HBx gene is involved in promoting the proliferation of HCC cells via up-regulating the expression of lncRNA-HEIH.