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

【摘要】： 目的研究KAI1對人鼻咽癌細(xì)胞體外增殖、侵襲及轉(zhuǎn)移能力等生物學(xué)行為的影響。 方法通過腺病毒介導(dǎo)將KAI1基因成功轉(zhuǎn)染到人鼻咽癌細(xì)胞株HNE-1中,應(yīng)用RT-PCR技術(shù)檢測轉(zhuǎn)染效果,采用四甲基偶氮唑藍(MTT)法測定鼻咽癌細(xì)胞轉(zhuǎn)染KAI1基因后體外增殖能力變化,采用流式細(xì)胞術(shù)測定細(xì)胞生長周期分布,采用Transwell侵襲小室檢測細(xì)胞侵襲能力改變,采用細(xì)胞同質(zhì)粘附實驗檢測細(xì)胞黏附力變化。 結(jié)果①當(dāng)rAd-KAI1病毒量為30MOI時,重組腺病毒對HNE-1細(xì)胞的轉(zhuǎn)染率達92%以上。②MTT法結(jié)果顯示,轉(zhuǎn)染后24h轉(zhuǎn)染組與對照組A值無顯著性差異(P0.05);轉(zhuǎn)染后48h、72h轉(zhuǎn)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(zhuǎn)染組G0/G1期細(xì)胞數(shù)量較對照組增多,S期細(xì)胞數(shù)量減少。④轉(zhuǎn)染組同質(zhì)黏附作用高于對照組(p0.05)。⑤侵襲實驗結(jié)果顯示,轉(zhuǎn)染組的穿膜細(xì)胞數(shù)為52.5±4.2,顯著低于對照組的167.4±3.5,差異有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論腺病毒成功介導(dǎo)抑癌基因KAI1轉(zhuǎn)染入鼻咽癌HNE-1細(xì)胞,并取得較高的轉(zhuǎn)染率。腫瘤轉(zhuǎn)移抑制基因KAI1對鼻咽癌細(xì)胞體外增殖能力具有抑制作用,并能將HNE-1細(xì)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(xì)胞生長周期從而抑制細(xì)胞增殖。轉(zhuǎn)染KAI1基因后HNE-1細(xì)胞侵襲力受到一定的抑制,且細(xì)胞同質(zhì)黏附力增強,提示:KAI1基因能夠抑制鼻咽癌HNE-1細(xì)胞侵襲能力,其機制可能為KAI1基因增強鼻咽癌細(xì)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(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R739.63

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