摘要
目的探讨肿瘤坏死因子-α诱导蛋白8样分子1(TIPE1)在食管癌中的表达及其作用机制。方法收集2018年1月-2019年1月陕西省安康市中医医院肿瘤外科手术切除食管癌10例,留取癌组织及癌旁正常组织备用。Westem-blot法检测TIPE1在食管癌组织及癌旁正常组织中的表达,利用脂质体将TIPE1空质粒及过表达载体转染到Eca109食管癌细胞,Western-blot法检测转染效果,CCK-8法检测细胞活力,流式细胞术检测细胞凋亡及细胞周期,Transwell法检测细胞侵袭能力,Westem-blot法检测细胞中细胞周期蛋白D1(CyclinD1)、CyclinB1、含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved-caspase 3)、cleaved-caspase 8、基质金属蛋白酶-2(MMP-2)、MMP-9表达。结果 TIPE1在食管癌组织中的表达量(0.13±0.01)显著低于癌旁正常组织(0.34±0.02),差异有统计学意义(t/P=29.698/0.000)。TIPE1过表达组TIPE1表达量(1.02±0.10)高于空白质粒组(0.21±0.02),差异有统计学意义(t/P=13.757/0.000)。TIPE1过表达组细胞活力(0.38±0.03)低于空白质粒组(0.48±0.04),差异有统计学意义(t/P=3.464/0.026)。TIPE1过表达组细胞早期凋亡率(16.43±1.65)%及晚期凋亡率(23.51±2.35)%均高于空白质粒组(2.58±0.27)%、(3.36±0.35)%,差异有统计学意义(t/P=14.348/0.000,14.689/0.000),且细胞周期主要阻滞于G1期(P<0.05)。TIPE1过表达组细胞侵袭数目[(42.36±4.24)个]少于空白质粒组[(158.32±14.33)个](t/P=13.440/0.000)。TIPE1过表达组CyclinD1、CyclinB1、cleaved-caspase 3、cleaved-caspase 8、MMP-2、MMP-9表达量均低于空白质粒组(t=10.457、10.922、52.034、46.540、11.585、13.555,P均=0.000)。结论 TIPE1在食管癌组织中低表达,TIPE1过表达能显著抑制Eca109食管癌细胞增殖与侵袭,并诱导细胞凋亡。
Objective To investigate the expression and mechanism of tumor necrosis factor a-inducible protein 8-like molecule 1(TIPE1) in esophageal cancer.Methods From January 2018 to January 2019,10 cases of esophageal cancer were surgically removed from the Department of Oncology,Ankang Hospital of Traditional Chinese Medicine,Shaanxi Province,and the cancer tissues and adjacent normal tissues were reserved.Western blot was used to detect the expression of TIPE1 in esophageal cancer tissues and adjacent normal tissues.TIPE1 empty plasmid and overexpression vector were transfected into Eca109 esophageal cancer cells using liposome.Western blot was used to detect the transfection effect.Cell viability was measured by the CCK 8 method.Flow cytometry was used to detect apoptosis and cell cycle.Transwell assay for cell invasiveness.Cytlin D1(CyclinD1),CyclinB1,cysteine-containing caspase caspase 3,cleaved caspase 8,matrix metalloproteinase 2(MMP-2),MMP 9 expression were detected by Western blot.Results The expression of TIPE1 in esophageal cancer tissues(0.13±0.01) was significantly lower than that in adjacent normal tissues(0.34±0.02),and the difference was statistically significant(t/P=29.698/0.000).The expression level of TIPE1 in TIPE1 overexpression group(1.02±0.10) was higher than that in blank plasmid group(0.21±0.02),and the difference was statistically significant(t/P=13.757/0.000).The cell viability of the TIPE1 overexpression group(0.38±0.03) was lower than that of the blank plasmid group(0.48±0.04),and the difference was statistically significant(t/P=3.464/0.026).The early apoptotic rate(16.43±1.65)% and the late apoptotic rate(23.51±2.35)% of TIPE1 overexpression group were higher than those of the blank plasmid group(2.58±0.27)%,(3.36±0.35)%,the difference was statistically significant.Significance(t/P=14.348/0.000,14.689/0.000),and the cell cycle was maialy blocked in the G1 phase(P <0.05).The number of cell invasions in the TIPE1 overexpression group [(42.36±4.24)] was less than that in the blank plasmid group [(158.32±14.33)](t/P=13.440/0.000).The expression levels of CyclinD1,CyclinB1,cleaved-caspase 3,cleaved caspase 8,MMP 2 and MMP 9 in TIPE1 overexpression group were lower than those in blank plasmid group(t=10.457,10.922,52.034,46.540,11.585,13.555,P=0.000).Conclusion TIPE1 is underexpressed in esophageal cancer tissues,and overexpression of TIPE1 can significantly inhibit the proliferation and invasion of Ecal09 esophageal cancer cells and induce apoptosis.
作者
王道军
赵山虎
夏平
吴应虎
WANG Daojun;ZHAO Shanhu;XIA Ping;WU Yinghu(Department of Oncology Surgery,Ankang Traditional Chinese Medicine Hospital,Shaanxi Province,Ankang 725000,China)
出处
《疑难病杂志》
CAS
2019年第12期1258-1262,共5页
Chinese Journal of Difficult and Complicated Cases
基金
国家自然科学基金资助项目(81702430)
陕西省卫生计生委科学技术基金项目(2018SF-244)
