KAI1对人鼻咽癌细胞株HNE-1增殖、侵袭及转移能力影响的实验研究

发布时间：2018-04-02 06:53

本文选题：KAI1　切入点：HNE-1　出处：《重庆医科大学》2010年硕士论文

【摘要】： 目的研究KAI1对人鼻咽癌细胞体外增殖、侵袭及转移能力等生物学行为的影响。方法通过腺病毒介导将KAI1基因成功转染到人鼻咽癌细胞株HNE-1中,应用RT-PCR技术检测转染效果,采用四甲基偶氮唑蓝(MTT)法测定鼻咽癌细胞转染KAI1基因后体外增殖能力变化,采用流式细胞术测定细胞生长周期分布,采用Transwell侵袭小室检测细胞侵袭能力改变,采用细胞同质粘附实验检测细胞黏附力变化。结果①当rAd-KAI1病毒量为30MOI时,重组腺病毒对HNE-1细胞的转染率达92%以上。②MTT法结果显示,转染后24h转染组与对照组A值无显著性差异(P0.05);转染后48h、72h转染组与对照组A值存在显著性差异(P0.05),其增殖抑制率分别为17%、30%。③转染组G0/G1期细胞数量较对照组增多,S期细胞数量减少。④转染组同质黏附作用高于对照组(p0.05)。⑤侵袭实验结果显示,转染组的穿膜细胞数为52.5±4.2,显著低于对照组的167.4±3.5,差异有统计学意义(P0.05)。结论腺病毒成功介导抑癌基因KAI1转染入鼻咽癌HNE-1细胞,并取得较高的转染率。肿瘤转移抑制基因KAI1对鼻咽癌细胞体外增殖能力具有抑制作用,并能将HNE-1细胞生长周期阻滞于G0/G1期,提示:KAI1基因可能通过阻滞鼻咽癌HNE-1细胞生长周期从而抑制细胞增殖。转染KAI1基因后HNE-1细胞侵袭力受到一定的抑制,且细胞同质黏附力增强,提示:KAI1基因能够抑制鼻咽癌HNE-1细胞侵袭能力,其机制可能为KAI1基因增强鼻咽癌细胞粘附力所致。
[Abstract]:Objective to study the effects of KAI1 on proliferation, invasion and metastasis of human nasopharyngeal carcinoma cells in vitro.Methods KAI1 gene was successfully transfected into human nasopharyngeal carcinoma (NPC) cell line HNE-1 by adenovirus-mediated transfection. RT-PCR technique was used to detect the transfection effect. The proliferation ability of nasopharyngeal carcinoma cell line transfected with KAI1 gene was measured by tetramethylazolyl tetrazolium (MTT) method in vitro.The cell growth cycle distribution was measured by flow cytometry, the change of cell invasion ability was detected by Transwell invasive chamber, and the change of cell adhesion force was detected by cell homogeneity adhesion assay.Results 1 when the amount of rAd-KAI1 virus was 30MOI, the transfection efficiency of recombinant adenovirus on HNE-1 cells was over 92%.There was no significant difference in A value between the transfection group and the control group 24 hours after transfection. There was a significant difference in A value between the transfection group and the control group at 48h after transfection, and the inhibition rate of proliferation was 170.303.The number of G0/G1 phase cells in the transfection group was more thin than that in the control group.The effect of homogenous adhesion in transfection group was higher than that in control group.The number of transmembrane cells in the transfection group was 52.5 卤4.2, which was significantly lower than that in the control group (167.4 卤3.5). The difference was statistically significant (P 0.05).Conclusion Adenovirus-mediated tumor suppressor gene KAI1 was successfully transfected into nasopharyngeal carcinoma HNE-1 cells and a high transfection rate was obtained.Tumor metastasis suppressor gene KAI1 can inhibit the proliferation of nasopharyngeal carcinoma cells in vitro and block the growth cycle of HNE-1 cells to G0/G1 phase, suggesting that the cell proliferation may be inhibited by blocking the growth cycle of nasopharyngeal carcinoma HNE-1 cells by blocking the growth cycle of HNE-1 cells.After transfection of KAI1 gene, the invasiveness of HNE-1 cells was inhibited to some extent, and the adhesion of HNE-1 cells was enhanced, suggesting that the invasion ability of nasopharyngeal carcinoma HNE-1 cells could be inhibited by 1: KAI1 gene, which may be caused by the enhancement of adhesion of nasopharyngeal carcinoma cells by KAI1 gene.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63

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