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| 肝癌患者锌指蛋白281的表达水平及其对肝癌细胞增殖凋亡的影响 |
| Expression level of zinc finger protein 281 in patients with liver cancer and its effects on proliferation and apoptosis of hepatoma carcinoma cells |
| 投稿时间：2018-07-09 |
| DOI：10.3969/j.issn.1000-0399.2019.06.004 |
| 中文关键词: 锌指蛋白281 肝癌细胞 增殖 凋亡 |
| 英文关键词: Zinc finger protein 281 Hepatoma carcinoma cell Proliferation Apoptosis |
| 基金项目:河南省高等学校重点科研项目计划（项目编号：16B320003） |
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| 中文摘要: |
| 目的 探讨肝癌患者锌指蛋白281（ZNF 281）的表达水平，并分析其对人肝癌细胞HepG2增殖和凋亡的影响。方法 收集河南大学第一附属医院2014年1月至2017年1月经手术切除后病理切片证实的肝癌标本82例及癌旁正常组织标本50例，采用免疫组化法检测肝癌和癌旁正常肝组织标本中ZNF 281蛋白表达情况，并分析ZNF 281阳性表达与临床病理参数[性别、年龄、肿瘤直径、国际恶性肿瘤分期(TNM分期)、淋巴结转移、甲胎蛋白(AFP)]的关系；采用荧光定量PCR检测上述组织中ZNF 281 mRNA的表达情况；采用沉默RNA （siRNA） ZNF 281转染人肝癌细胞HepG2，根据转染情况分为空白对照组（正常培养的HepG2）、阴性对照组、siRNA ZNF 281组[转染ZNF 281基因特异性siRNA干扰组]，并用荧光定量PCR检测转染后HepG2细胞中ZNF 281 mRNA的表达，四甲基偶氮唑蓝（MTT）法检测转染后细胞增殖活性，流式细胞仪检测细胞凋亡率。结果 肝癌组织中ZNF 281 mRNA的相对表达量高于正常肝组织（P<0.05）；肝癌组织ZNF 281蛋白阳性表达率为74.39%，高于癌旁正常肝组织的18.0%（P<0.05），且ZNF 281蛋白阳性表达患者肿瘤直径>5 cm，TNM为Ⅲ~Ⅳ期，淋巴结转移比例分别为65.57%、73.77%、45.90%，高于ZNF 281蛋白表达阴性的38.10%、38.10%、14.29%（P<0.05）。siRNA ZNF 281组HepG2细胞中ZNF 281 mRAN表达量低于空白对照组和阴性对照组（P<0.05）；siRNA ZNF 281组细胞生长抑制率和细胞凋亡率均高于空白对照组和阴性对照组（P<0.05）；空白对照组和阴性对照组HepG2细胞中ZNF 281 mRAN表达量及细胞生长抑制率和细胞凋亡率比较，差异无统计学意义（P>0.05）。结论 肝癌组织中ZNF 281蛋白及其RNA均呈高表达；沉默ZNF 281基因可抑制人肝癌细胞HepG2增殖，诱导其凋亡。 |
| 英文摘要: |
| Objective To explore the expression level of zinc finger protein 281 (ZNF 281) in patients with liver cancer, and analyze its effects on proliferation and apoptosis of hepatoma carcinoma cell HepG2. Methods A total of 82 liver cancer specimens and 50 adjacent normal tissue specimens confirmed by pathological sections after surgical excision in the hospital from January 2014 to January 2017 were collected. The expressions of ZNF 281 protein in liver cancer and adjacent normal tissues were detected by immunohistochemistry. The relationship between positive expression of ZNF 281 protein and clinicopathological parameters[gender, age, tumor diameter, international malignant tumor stage (TNM stage), lymph node metastasis, alpha-fetoprotein (AFP)] was analyzed. The expression of ZNF 281 mRNA in the above tissues was detected by fluorescent quantitative PCR. The human hepatoma carcinoma cell HepG2 was transfected by (small interfering RNA) siRNA ZNF 281. According to the condition of transfection, the specimens were divided into blank control group (normal cultured HepG2), negative control group[transfected with ZNF 281 gene non-specific negative silencing RNA (siRNA) interference group] and siRNA ZNF 281 group[transfected with ZNF 281 gene specific siRNA interference group]. The expression of ZNF 281 mRNA in HepG2 cells after transfection was detected by real-time PCR. The proliferation activity of the cells after transfection was detected by methyl thiazolyl tetrazolium (MTT) and the apoptotic rate measured by flow cytometry. Results The relative expression of ZNF 281 mRNA in liver cancer tissues was higher than that in normal liver tissues (P<0.05). The expression rate of ZNF 281 protein in liver cancer tissues was higher than that in normal adjacent liver tissue (74.39% vs 18.0%, P<0.05). The rate of tumor diameter greater than 5 cm, TNM in Ⅲ to IV stage and lymph node metastasis in patients with ZNF 281 protein positive expression was 65.57%, 73.77%, and 45.90%, respectively, which was higher than that in patients with ZNF 281 protein negative expression (38.10%, 38.10%, and 14.29%) (P<0.05). The ZNF 281 mRAN expression in HepG2 cells of siRNA ZNF 281 group was lower than that of blank control group and negative control group (P<0.05). The cell growth inhibitory rate and apoptotic rate of hepatoma cells in siRNA ZNF 281 group were higher than those in blank control group and negative control group (P<0.05). There was no significant difference between blank control group and negative control group in ZNF 281 mRAN expression in HepG2 cells or cell growth inhibitory rate and apoptotic rate (P>0.05). Conclusions ZNF 281 protein and mRNA are highly expressed in liver cancer tissues. Silencing ZNF 281 gene may inhibit the proliferation of human hepatoma carcinoma cell HepG2 and induce apoptosis. |
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