本文選題：KAI1　切入點：HNE-1　出處：《重慶醫(yī)科大學》2010年碩士論文

【摘要】： 目的研究KAI1對人鼻咽癌細胞體外增殖、侵襲及轉移能力等生物學行為的影響。 方法通過腺病毒介導將KAI1基因成功轉染到人鼻咽癌細胞株HNE-1中,應用RT-PCR技術檢測轉染效果,采用四甲基偶氮唑藍(MTT)法測定鼻咽癌細胞轉染KAI1基因后體外增殖能力變化,采用流式細胞術測定細胞生長周期分布,采用Transwell侵襲小室檢測細胞侵襲能力改變,采用細胞同質粘附實驗檢測細胞黏附力變化。 結果①當rAd-KAI1病毒量為30MOI時,重組腺病毒對HNE-1細胞的轉染率達92%以上。②MTT法結果顯示,轉染后24h轉染組與對照組A值無顯著性差異(P0.05);轉染后48h、72h轉染組與對照組A值存在顯著性差異(P0.05),其增殖抑制率分別為17%、30%。③轉染組G0/G1期細胞數量較對照組增多,S期細胞數量減少。④轉染組同質黏附作用高于對照組(p0.05)。⑤侵襲實驗結果顯示,轉染組的穿膜細胞數為52.5±4.2,顯著低于對照組的167.4±3.5,差異有統(tǒng)計學意義(P0.05)。 結論腺病毒成功介導抑癌基因KAI1轉染入鼻咽癌HNE-1細胞,并取得較高的轉染率。腫瘤轉移抑制基因KAI1對鼻咽癌細胞體外增殖能力具有抑制作用,并能將HNE-1細胞生長周期阻滯于G0/G1期,提示:KAI1基因可能通過阻滯鼻咽癌HNE-1細胞生長周期從而抑制細胞增殖。轉染KAI1基因后HNE-1細胞侵襲力受到一定的抑制,且細胞同質黏附力增強,提示:KAI1基因能夠抑制鼻咽癌HNE-1細胞侵襲能力,其機制可能為KAI1基因增強鼻咽癌細胞粘附力所致。
[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 鹵4.2, which was significantly lower than that in the control group (167.4 鹵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.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R739.63

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