IL-6/JAK2/STAT3对口腔鳞状细胞癌生物学行为的影响
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1.中南大学湘雅三医院;2.湖南省肿瘤医院/中南大学湘雅医学院附属肿瘤医院;3.中南大学生命科学学院;4.湖南省肿瘤医院//中南大学湘雅医学院附属肿瘤医院

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湖南省自然科学基金(2019JJ40216)


Relationship between IL-6 / JAK2 / STAT3 and the biological behavior of oral squamous cell carcinoma
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1.Department of Stomatology,Third Xiangya Hospital,Central South University,Changsha,Hunan;2.Head and Neck Surgery Department,Hunan Cancer Hospital / Affiliated Cancer Hospital,Xiangya Medical College,Central South University

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    摘要:

    目的 观察IL-6/JAK2/STAT3对口腔鳞状细胞癌细胞生物学行为的影响。方法 对口腔鳞状细胞癌细胞系进行慢病毒转染敲低IL-6基因,设立两组对照,一组未干预组,另一组为空白载体组,通过CCK8实验和克隆形成实验检测IL-6对口腔鳞状细胞癌细胞增殖情况的影响,通过流式细胞术检测细胞周期变化,通过Transwell实验检测IL-6对口腔鳞癌细胞侵袭和迁移能力的影响。利用实时荧光定量PCR以及Western Blot方法检测口腔鳞癌细胞中IL-6基因表达下调后其下游通路基因p-STAT3、p-JAK2、与肿瘤血管新生有关的蛋白VEGFA、细胞周期调控蛋白Cyclin B1、细胞基质沉积相关蛋白MMP-2、细胞凋亡抑制蛋白Survivin、上皮向间质转化相关的蛋白Snail和E-Cadherin的表达情况。结果 敲低IL-6的口腔鳞癌细胞与对照组(方法中未见交代对照组,结果中怎么出现呢?请前后一致!!)细胞比较,增殖率受到明显抑制,差异有统计学意义( F值=9.09/10.08,P=0.015/0.011,P<0.05);敲低IL-6的口腔鳞癌细胞中G1期细胞百分比下降,G2期细胞百分比上升,差异具有统计学意义(F值=5.53,P=0.044,P<0.05);敲低IL-6的口腔鳞癌细胞与对照细胞比较,侵袭能力和迁移能力明显下降,差异有统计学意义(F值=23.59/53.11,P=0.000/0.000,P<0.01);口腔鳞癌细胞IL-6表达下调后,JAK2、STAT3(F值=45.30/34.58,P=0.000/0.001)、VEGFA(F值=28.18/43.41,P=0.001/0.000)、Cyclin B1(F值=18.11/5.41,P=0.003/0.045)、MMP-2(F值=66.91/18.47,P=0.000/0.03)、Snai(F值=1.91/10.88,P=0.03/0.01)l表达随之明显下降,而E-Cadherin(F值=176.92/97.17,P=0.000/0.000)表达显著增加,差异有统计学意义(P<0.05)结论 IL-6基因敲低后可明显抑制口腔鳞癌细胞的生长与增殖能力,可能诱导细胞G2期阻滞;抑制STAT3、JAK2、VEGFA、Cyclin B1、MMP-2、Snail和Survivin表达,促进E-Cadherin蛋白表达。

    Abstract:

    Objective To investigate the effects of IL-6 /JAK2 /STAT3 on the biological behavior of oral squamous cancer cells.Methods The oral squamous cell carcinoma cell line was transfected with lentivirus, and the IL-6 gene was knockout.The effect of IL-6 on the proliferation of oral squamous cell carcinoma cells was detected by CCK8 experiment and clonal formation experiment, the cellular cycle change was detected by flow cytometry, and the influences of IL-6 on invasion and migration ability of oral squamous cell carcinoma cells were detected by Transwell experiment.Then real-time fluorescence quantitative PCR and Western blot method were used to detect the expression of the downstream channel gene STAT3, JAK2, protein VEGFA related to tumor angiogenesis, cell cycle regulatory protein Cyclin B1, cell matrix deposition-related protein MMP-2, apoptosis suppressor protein Survivin, and protein Snail and E-Cadherin related to epithelial-to-interstitial transformation in oral squamous cell carcinoma cells after downregulation of IL-6 gene expression.Results Compared with the control group, the oral squamous cell carcinoma cells with knockout IL-6 was significantly inhibited in proliferation rate, and the difference was statistically significant (P<0.05); The percentage of G1-phase cells in oral squamous cell carcinoma cells with knockout IL-6 decreased significantly, and the percentage of G2-phase cells increased significantly, and the difference was statistically significant(P<0.05). The invasive ability and migration ability of oral squamous cell carcinoma cells with knockout IL-6 decreased significantly compared with control cells, and the difference was statistically significant(P<0.05); Compared with the control group, the expression of STAT3, JAK2, VEGFA, Cyclin B1, MMP-2, Snail and Survivin protein decreased significantly while E-Cadherin protein expression increased apparently in oral squamous cell carcinoma cells with knockout IL-6 (P<0.05).Conclusion IL-6/JAK2/STAT3 signaling pathway can promote proliferation and migration of oral squamous cell carcinoma.

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  • 收稿日期:2021-12-03
  • 最后修改日期:2022-01-08
  • 录用日期:2022-01-11
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