摘要
目的探讨三联组氨酸核苷酸结合蛋白1在食管癌中的表达及机制.方法选择Eca-109食道癌细胞株实验室细胞培养,用pLVX-IRES-ZsCreen1慢病毒表达Hint1载体,感染293T细胞纯化浓缩后,感染Eca-109细胞得到Eca-109^Hint1+组细胞;测定两组细胞绿色荧光蛋白的表达,检测转染是否成功;用蛋白质印迹法(Western blot)法检测细胞Hint1蛋白的表达;用Transwell侵袭实验检测不同组细胞的侵袭能力;用流式细胞仪检测细胞周期;用噻唑蓝(MTT)法检测细胞生长情况.结果Eca-109Hint1+组细胞中荧光蛋白平均表达率(68.26±3.12)%,而对照组细胞未见绿色荧光表达(t=8.250,P<0.01);Western blot法检测Eca-109^Hint1+组细胞中Hint1表达为(1.86±0.02),而Eca-109组细胞中Hint1表达为(0.61±0.01,t=5.529,P<0.05);Eca-109^Hint1+组细胞的侵袭能力为(49.36±3.02),而Eca-109组细胞侵袭能力为(114.11±10.01),Eca-109^Hint1+组细胞的侵袭能力低于Eca-109组细胞的侵袭能力(t=8.988,P<0.05);Eca-109组细胞细胞周期为(50.31±0.41),Eca-109^Hint1+组细胞的细胞周期为(41.24±0.62);MTT实验显示Eca-109^Hint1+组细胞的吸光度(A)值为(0.82±0.01),Eca-109组细胞的A值为(0.57±0.02),Eca-109^Hint1+组细胞的A值高于Eca-109组细胞的A值(t=6.527,P<0.05).结论三联组氨酸核苷酸结合蛋白1在食管癌中的表达时,细胞周期变长,细胞活力增加,对于食管癌细胞具有抑制作用.
Objective To study the expression and mechanism of trihistidine nucleotide binding protein 1 in esophageal carcinoma.Methods The Eca-109 esophageal cancer cell line was cultured in laboratory.The Hint1 vector was expressed by pLVX-IRES-ZsCreen1 lentivirus.After 293T cells were purified and concentrated,the Eca-109 cells were infected to obtain Eca-109 Hint1 group cells.To detect the expression of green fluorescent protein(CFP)in two groups,to detect whether the transfection was successful or not,to detect the expression of Hint1 protein by Western blotting method,detection of invasiveness in different groups of cells by Transwell invasion assay,to detect the cell cycle by flow cytometry,and to detect the cell growth by methyl thiazol tetrazolium(MTT)method.Results The average expression rate of fluorescent protein in Eca-109 Hint1 group was(68.26±3.12)%(t=8.250,P<0.01).These results indicated that the transfection of human esophageal cancer cells infected with Hint1 vector virus was successful.The expression of Hint1 in Eca-109 Hint1 group was(1.86±0.02),and that of Hint1 in Eca-109 group was(0.61±0.01)by Western blot assay(t=5.529,P<0.05).The invasiveness of Eca-109^Hint1+group was(49.36±3.02),while that of Eca-109 group was(114.11±10.01).The invasiveness of Eca-109^Hint1+group was significantly lower than that of Eca-109 group(t=8.988,P<0.05).The cell cycle of Eca-109 group was(50.31±0.41),and that of Eca-109 Hint1 group was(41.24±0.62).On the third day of culture,the absorbance of cells in Eca-109 Hint1 group was(0.82±0.01)and that in Eca-109 group was(0.57±0.02).The results showed that the absorbance value of Eca-109 Hint1 group was significantly higher than that of Eca-109 group(t=6.527,P<0.05).Conclusion The expression of tri-histidine nucleotides binding protein-1 in esophageal carcinoma resulted in the increase of cell cycle and cell viability,which could inhibit the growth of esophageal cancer cells.
作者
吕俊杰
陈玉兄
张振中
吴旭
Lv Junjie;Chen Yuxiong;Zhang Zhenzhong;Wu Xu(Department of Thoracic Surgery Huaian First Hospital Affiliated to Nanjing Medical University,Huaian 223200,China;Department of Thoracic,Nanfang Hospital Southern Medical Vniversity,Guangzhou 510155,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第9期1551-1553,共3页
Chinese Journal of Experimental Surgery
关键词
三联组氨酸核苷酸结合蛋白1
食管癌
细胞表达及抑制
Trihistidine nucleotide binding protein 1
Esophageal carcinoma
Cell expression and inhibition
