论文部分内容阅读
目的观察幽门螺杆菌(Hp)野生株与iceA基因插入失活突变株对人胃上皮细胞株SGC7901细胞生长的影响,研究Hp毒力基因iceA相关的细胞毒性效应。方法分别将幽门螺杆菌野生株与iceA基因插入失活突变株与SGC7901细胞共培养48h,观察比较细胞形态学变化;用MTT法检测分析共培养12～36h细胞增殖活性;流式细胞仪分析48h细胞周期和24、48h细胞凋亡。结果 Hp野生株组和突变株组引起相似的细胞形态学改变;30h和36h时各组细胞的增殖活性表现出明显差异(P<0.05),培养30h时突变株组(0.745±0.006)的细胞增殖活性高于野生株组(0.728±0.017)(P<0.05);细胞周期结果显示,野生株组G2期阻滞,而突变株组为S期阻滞;培养24、48h野生株组[(33.04±10.69)%、(52.28±3.41)%]和突变株组[(44.23±11.34)%、(57.32±0.88)%]均可以诱导细胞产生明显凋亡(P<0.05),但二者在诱导细胞凋亡的能力方面差异无统计学意义(P>0.05)。结论幽门螺杆菌对于SGC7901细胞生长状态产生明显的影响,iceA基因的插入失活突变对于SGC细胞的形态变化和凋亡的影响无明显差异,对细胞增殖和细胞周期影响差异较明显。
Objective To observe the effects of wild-type Hp and iceA gene inactivation on the growth of human gastric epithelial cell line SGC7901, and to study the cytotoxicity effect of iceA related to Hp virulence gene. Methods Mutant Helicobacter pylori and iceA gene inactivated mutant and SGC7901 cells were co-cultured for 48h respectively. Morphological changes of cells were observed and compared. MTT assay was used to detect the proliferative activity of cells cultured for 12h to 36h. Flow cytometry analysis was performed for 48h Cell cycle and 24,48h apoptosis. Results The Hp wild-type group and the mutant group caused similar cell morphological changes. The proliferation activity of each group was significantly different at 30h and 36h (P <0.05). After 30h incubation, the cells in the mutant group (0.745 ± 0.006) (P <0.05). The results of cell cycle showed that the G2 phase of the wild-type group was blocked, while the mutant group was S-phase arrest. The 24 and 48-h wild-type strain [ 33.04 ± 10.69%, 52.28 ± 3.41%, and 44.23 ± 11.34%, 57.32 ± 0.88%, respectively) were significantly higher than those in the control group (P <0.05) There was no significant difference in the ability to induce apoptosis (P> 0.05). Conclusion Helicobacter pylori has a significant effect on the growth of SGC7901 cells. There is no significant difference in the morphological changes and apoptosis of SGC7901 cells induced by insertion of iceA gene, and the effect on cell proliferation and cell cycle is obvious.