Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
Department of Breast Surgery, Renmin Hospital of Shiyan, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China.
Mol Med Rep. 2023 May;27(5). doi: 10.3892/mmr.2023.12985. Epub 2023 Mar 24.
Hepatitis B virus (HBV) integration into human genome causes hepatocellular carcinoma (HCC). The present study used inverse nested PCR; the full sequence of HBV DNA fragments of the chrX: 111009033 integration site was detected (987 bp), containing two fragments of double‑stranded linear DNA with the same orientation (1,744‑1,094 and 1,565‑1,228 nt). By reverse transcription‑quantitative PCR, HBV‑cell fusion transcript was observed in HepG2.2.15 cells. The mean copy number of this site in cells with HO treatment (8.73x10±1.65x10 copies/cell) was significantly higher than that in the cells without HO treatment (3.02x10±2.33x10 copies/cell; P<0.0001). The mean levels of P21‑activated kinase 3 (PAK3) were 15.67±5.65 copies/cell in HepG2.2.15 cells with HO treatment, significantly higher than in the cells without HO treatment (11.34±4.58 copies/cell, P=0.0076) and in HepG2 cells (5.92±1.54 copies/cell, P<0.0001). Significant difference of PAK3 levels was also found between HepG2.2.15 cells without HO treatment and HepG2 cells (11.34±4.58 vs. 5.92±1.54 copies/cell, P<0.0001). The average copy numbers of the integration site chrX: 111009033 were positively correlated with the average levels of PAK3 (P=0.0013). The overall trend of PAK3 expression was significantly increased in HepG2.2.15 cells with HO treatment compared with that in HepG2.2.15 cells without HO treatment (37.63±8.16 and 31.38±7.94, P=0.008) and HepG2 cells (21.67±7.88, P<0.0001). In summary, the chrX: 11009033 integration site may originate from primary human hepatocytes, occurrence and clonal expansion of which may upregulate PAK3 expression, which may contribute to hepatocarcinogenesis.
乙型肝炎病毒（HBV）整合到人类基因组中会导致肝细胞癌（HCC）。本研究采用反向巢式 PCR；检测到整合位点 chrX:111009033 的 HBV DNA 片段全长序列（987bp），包含两个相同方向的双链线性 DNA 片段（1,744-1,094 和 1,565-1,228nt）。通过逆转录定量 PCR，在 HepG2.2.15 细胞中观察到 HBV-细胞融合转录本。HO 处理组该位点的平均拷贝数（8.73x10±1.65x10 拷贝/细胞）明显高于未处理组（3.02x10±2.33x10 拷贝/细胞；P<0.0001）。HO 处理组 HepG2.2.15 细胞中 P21 激活激酶 3（PAK3）的平均水平为 15.67±5.65 拷贝/细胞，明显高于未处理组（11.34±4.58 拷贝/细胞，P=0.0076）和 HepG2 细胞（5.92±1.54 拷贝/细胞，P<0.0001）。HO 处理组 HepG2.2.15 细胞与 HepG2 细胞之间的 PAK3 水平也存在显著差异（11.34±4.58 vs. 5.92±1.54 拷贝/细胞，P<0.0001）。整合位点 chrX:111009033 的平均拷贝数与 PAK3 的平均水平呈正相关（P=0.0013）。与未用 HO 处理的 HepG2.2.15 细胞相比，用 HO 处理的 HepG2.2.15 细胞中 PAK3 的表达呈总体上升趋势（37.63±8.16 和 31.38±7.94，P=0.008）和 HepG2 细胞（21.67±7.88，P<0.0001）。综上所述，chrX:11009033 整合位点可能来源于原代人肝细胞，其发生和克隆扩增可能上调 PAK3 的表达，这可能有助于肝癌的发生。
