Hfgl2在宫颈癌患者组织中的表达及其临床意义

Expression of Hfgl2 in patients with cervical cancer and its clinical significance

摘要

目的 分析人纤维介素(Hfgl2)在宫颈癌组织中的表达及其意义,探究其对宫颈癌恶性生物学行为的影响。方法 选取2016年1月至2018年1月在榆林市第一医院经病理确诊的50例宫颈癌患者的癌组织标本作为实验组,另外收集同期50例正常宫颈组织作为对照组.采用免疫组化和逆转录聚合酶链反应(RT-PCR)检测组织,蛋白质印迹法(WB)检测细胞中Hfgl2的表达,分析其与宫颈癌临床病理特征的关系;采用基因敲降下调Hfgl2表达,四甲基偶氮唑盐比色法(MTT)检测细胞增殖能力,WB方法检测血管生成及细胞增殖、迁移相关蛋白的表达。结果 Hfgl2在宫颈癌组织中的阳性表达率高于正常宫颈组织,差异有统计学意义(χ²=73.990,P<0.05);Hfgl2在宫颈癌组织的免疫组化评分高于正常宫颈组织,差异有统计学意义(t=8.022,P<0.05);RT-PCR测定表明,实验组Hfgl2 mRNA阳性表达率高于对照组,差异有统计学意义(χ²=61.455,P<0.05);Hfgl2在宫颈癌国际妇产科联盟(FIGO)分期Ⅲ-Ⅳ期中的阳性表达率高于Ⅰ期、Ⅱ期组,在病理分级G2-G3级宫颈癌组的阳性表达率高于G1级组,在有淋巴转移的宫颈癌组织中阳性表达率高于无淋巴结转移组,差异均有统计学意义(χ²值分别为6.737、7.147、12.037,P<0.05);Hfgl2在宫颈癌细胞系中均高表达,敲降Hfgl2后,细胞体外增殖能力明显降低,人宫颈癌细胞Hela血管生成及增殖迁移相关蛋白如血管内皮生长因子受体1(VEGFR1)、蛋白激酶B(PKB/Akt)、磷脂酶C(PLC)-γ、基质金属蛋白酶(MMP)2和MMP9表达均明显下调,差异有统计学意义(t值介于4.904~8.304之间,P<0.05)。结论 Hfgl2在宫颈癌组织中高表达且可能促进肿瘤血管生成及浸润生长,有望成为宫颈癌靶向治疗新方向.

Abstract

Objective To analyze the expression and significance of human fibrinogen-like protein (Hfgl2) in cervical cancer tissues and explore its impact on the malignant biological behavior of cervical cancer. Methods We selected tumor specimens from 50 cervical cancer patients diagnosed pathologically at the First Hospital of Yulin from January 2016 to January 2018 as the experimental group, and collected 50 normal cervical tissue specimens from the same period as the control group. Immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect tissue samples. Western blot (WB) was employed to assess Hfgl2 expression in cells, and the relationship with clinical pathological characteristics of cervical cancer was analyzed. Gene knockdown was utilized to downregulate Hfgl2 expression, and the methyl thiazolyl diphenyl-tetrazolium bromide (MTT) assay was employed to measure cell proliferation ability, with WB used to evaluate the expression of proteins related to angiogenesis, cell proliferation, and cell migration. Results The positive expression rate of Hfgl2 in cervical cancer tissues was higher than that in normal cervical tissues, with statistically significant differences (χ²=73.990, P<0.05); The immunohistochemical score of Hfgl2 in cervical cancer tissues was higher than those in normal cervical tissues, with statistically significant differences (t=8.022, P<0.05); RT-PCR analysis revealed a higher positive rate of Hfgl2 mRNA expression in the experimental group compared to the control group, with statistically significant differences (χ²=61.455, P<0.05); The positive expression rate of Hfgl2 in Federation International of Gynecology and Obstetrics (FIGO) stage Ⅲ-Ⅳ cervical cancer was higher than in stage I and II groups, higher in pathological grade G2-G3 cervical cancer than in G1, and higher in cervical cancer tissues with lymph node metastasis than in those without, all with statistically significant differences (χ²=6.737, 7.147, and 12.037 respectively, P<0.05); Hfgl2 was highly expressed in cervical cancer cell lines, and knockdown of Hfgl2 resulted in a significant decrease in cell vitro proliferation ability, and the expression of angiogenesis and proliferation-related proteins vascular endothelial growth factor receptor 1 (VEGFR1), protein kinase B (PKB/Akt), phospholipase C (PLC)-γ, matrix metalloproteinase (MMP) 2, and MMP9 in human cervical cancer cells HeLa were significantly downregulated, with statistically significant differences (t=4.904-8.304, P<0.05). Conclusion Hfgl2 is highly expressed in cervical cancer tissue and may promote tumor angiogenesis and invasive growth, which is expected to become a new direction for targeted treatment in cervical cancer.

关键词

人纤维介素 / 宫颈癌 / 临床病理特征 / 血管生成 / 增殖迁移

Key words

human fibrinogen-like protein 2 / cervical cancer / clinical pathological characteristics / angiogenesis / proliferative and migration

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基金资助

国家自然科学基金项目(82203465)