本文選題：�？颂婺� + 肺癌　； 參考：《川北醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:初步探討鹽酸�？颂婺釋Ψ伟┘�(xì)胞的增殖抑制作用與EGFR基因突變狀態(tài)的關(guān)系,為臨床上細(xì)胞藥敏試驗(yàn)作為EGFR-TKIs療效預(yù)測方法之一提供參考。方法:以體外培養(yǎng)的A549和HCC827肺癌細(xì)胞株為實(shí)驗(yàn)對象,通過形態(tài)學(xué)觀察和免疫細(xì)胞化學(xué)染色法鑒定兩細(xì)胞株,采用x TAG液相芯片技術(shù)檢測細(xì)胞株EGFR基因突變狀態(tài),CCK-8法檢測�？颂婺釋杉�(xì)胞株的增殖抑制作用,Annexin V-FITC/PI雙染法檢測�？颂婺釋杉�(xì)胞株的促凋亡作用,實(shí)驗(yàn)數(shù)據(jù)應(yīng)用SPSS21.0軟件進(jìn)行統(tǒng)計(jì)學(xué)分析,檢驗(yàn)水準(zhǔn)為P0.05。結(jié)果:1)兩細(xì)胞株形態(tài)學(xué)表現(xiàn)和免疫細(xì)胞化學(xué)染色情況均符合A549和HCC827肺癌細(xì)胞株的特性。2)EGFR基因突變狀態(tài):A549細(xì)胞株為野生型,HCC827細(xì)胞株為19外顯子缺失突變型(p.E746_A750del)。3)�？颂婺釋549細(xì)胞株作用48h的半數(shù)抑制濃度(IC50)為9.65±0.78μmol/L,對HCC827細(xì)胞株作用的IC50為0.12±0.01μmol/L,兩者比較差異有統(tǒng)計(jì)學(xué)意義(P=0.000)。4)選用0.1μmol/L鹽酸�？颂婺嶙饔肁549和HCC827細(xì)胞株48h,兩者細(xì)胞抑制率分別為0和(45.31±1.33)%(P=0.000),細(xì)胞凋亡率分別為(1.81±0.47)%和(62.25±2.45)%(P=0.000);選用1μmol/L鹽酸�？颂婺嶙饔肁549和HCC827細(xì)胞株48h,兩者細(xì)胞抑制率分別為(18.95±0.78)%和(75.53±2.98)%(P=0.000),兩者細(xì)胞凋亡率分別為(39.91±2.04)%和(89.26±3.81)%(P=0.000)。結(jié)論:1)本課題初步證實(shí)了免疫細(xì)胞化學(xué)染色鑒定A549和HCC827肺癌細(xì)胞株的可行性,研究中的免疫細(xì)胞化學(xué)染色結(jié)果可能成為其他研究中鑒定該兩細(xì)胞株時的參考依據(jù);2)課題中�？颂婺釋GFR基因突變型的HCC827肺腺癌細(xì)胞株的增殖抑制作用明顯強(qiáng)于EGFR基因野生型的A549肺癌細(xì)胞株,提示研究方法真實(shí)、可靠,可為臨床上EGFR-TKIs細(xì)胞藥敏試驗(yàn)的實(shí)施提供一定參考。
[Abstract]:Objective: to investigate the relationship between the inhibitory effect of Ektinib hydrochloride on the proliferation of lung cancer cells and the mutation status of EGFR gene, and to provide a reference for clinical chemosensitivity test as one of the methods for predicting the curative effect of EGFR-TKIs. Methods: A549 and HCC827 lung cancer cell lines were cultured in vitro and identified by morphological observation and immunocytochemical staining. X TAG liquid chip technique was used to detect the mutation status of EGFR gene in cell line and CCK-8 method was used to detect the inhibitory effect of Ectini on the proliferation of two cell lines. Annexin V-FITC/PI double staining was used to detect the apoptosis-promoting effect of Ectini on the two cell lines. The experimental data were analyzed by SPSS21.0 software, and the test level was P0.05. Results the morphological features and immunocytochemical staining of the two cell lines were consistent with the characteristics of A549 and HCC827 lung cancer cell lines. IC50 was 9.65 鹵0.78 渭 mol / L in A549 cell line and 0.12 鹵0.01 渭 mol / L in HCC827 cell line for 48 h. The difference was statistically significant (P < 0.05). The inhibition rate of A549 cell line and HCC827 cell line treated with 0.1 渭 mol/L epetinib hydrochloride for 48 h was significant. The cell apoptosis rates were 1.81 鹵0.47% and 62.25 鹵2.45%, respectively, and the inhibitory rates of 1 渭 mol/L hydrochloride on A549 and HCC827 cells for 48 h were 18.95 鹵0.78% and 75.53 鹵2.98%, respectively, and the apoptotic rates were 39.91 鹵2.04% and 89.26 鹵3.81%, respectively. Conclusion 1) the feasibility of identification of A549 and HCC827 lung cancer cell lines by immunocytochemical staining was preliminarily confirmed. The results of immunocytochemical staining in the study may be the reference basis for identifying the two cell lines in other studies. The inhibitory effect of Ectini on the proliferation of EGFR mutant HCC827 lung adenocarcinoma cell lines is significantly stronger than that of other studies. EGFR gene wild-type lung cancer cell line A549, The results suggest that the research method is true and reliable, and can provide some reference for the clinical application of EGFR-TKIs cell drug sensitivity test.
【學(xué)位授予單位】：川北醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R734.2

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