本文選題：肺腺癌　切入點：GPC3　出處：《重慶醫(yī)科大學(xué)》2016年碩士論文

【摘要】：目的:研究用靶向GPC3基因的si RNA干擾人肺腺癌A549細胞中GPC3基因的表達后,對A549細胞生物學(xué)行為產(chǎn)生的影響,說明GPC3基因在A549細胞株中的作用,進而為人類肺腺癌的早期診斷、預(yù)測預(yù)后及治療手段提供理論基礎(chǔ)。方法:將細胞接種于不含抗生素的培養(yǎng)基中,24h后用Opti-MEM稀釋Lipofectamine 2000和siRNA的混合液,再加入到需要轉(zhuǎn)染的細胞中。37℃培養(yǎng)24h后,通過western blot和熒光定PCR檢測空白組,NC組及干擾組的細胞GPC3蛋白/基因表達量。通過CCK-8實驗、劃痕實驗以及流式細胞術(shù)檢測干擾了GPC3基因之后的A549細胞的生物學(xué)行為的變化。結(jié)果:干擾組GPC 3基因的蛋白及核算表達水平明顯低于空白組和NC組,差異具有顯著性(P0.05);而NC組和空白組的表達量沒有顯著差異。與NC組和空白組A549細胞相比,干擾組的A549細胞遷移和增殖能力明顯降低,而細胞凋亡率明顯增加(P0.01)。結(jié)論:利用Lipofectamine 2000介導(dǎo)的基因轉(zhuǎn)染技術(shù)成功的干擾了A549細胞中GPC3基因的表達。當(dāng)GPC3的表達降低時,A549細胞的增殖和遷移能力都減低;相反,A549細胞的凋亡率增加。
[Abstract]:Objective: To study the expression of GPC3 targeting GPC3 gene by Si interference of RNA human lung adenocarcinoma cell A549 gene, the influence on the biological behavior of A549 cells, indicating that the effect of GPC3 gene in A549 cell line, and then for the early diagnosis of human lung adenocarcinoma, and provide a theoretical basis for prognosis and treatment. Methods: the cells were seeded in culture medium without antibiotics, 24h with a mixture of Opti-MEM diluted Lipofectamine 2000 and siRNA, and then added to the 24h culture.37 C need transfected cells after western and blot by fluorescence quantitative PCR detection of the blank group, the expression of NC group and interference group of cells GPC3 protein / gene the amount of CCK-8. Through the experiment, changes of scratch test and flow cytometry interfere with the biological behavior of GPC3 gene in A549 cells. Results: the protein and accounting interference group GPC 3 gene expression levels were significantly lower than the blank group and In NC group, the difference was significant (P0.05); and the expression of NC group and blank group was no significant difference. Compared with the NC and A549 cells in the blank group, A549 cell migration and proliferation ability of interference group was significantly decreased, and the apoptosis rate increased significantly (P0.01). Conclusion: the use of Lipofectamine 2000 mediated gene transfection technique successfully interfered with the expression of GPC3 gene in A549 cells. The decreased expression of GPC3, A549 cell proliferation and migration are reduced; on the contrary, the apoptosis rate of A549 cells increased.

【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】：2016
【分類號】：R734.2

【參考文獻】

相關(guān)期刊論文 前1條

1 董志偉,喬友林,李連弟,陳育德,王潤田,雷通海,饒克勤,王汝寬,趙平,游偉程,魯鳳珠,戴旭東,王國清,羅賢懋,周海城;中國癌癥控制策略研究報告[J];中國腫瘤;2002年05期

，

本文編號：1704545