本文選題：肺腺癌 + VEGF-A��； 參考：《安徽醫(yī)科大學(xué)》2016年碩士論文

【摘要】：目的探討轉(zhuǎn)染mi RNA-16后作用不同時間對體外培養(yǎng)的人肺腺癌A549細(xì)胞增殖的影響,以及對肺腺癌A549細(xì)胞內(nèi)VEGF-A表達(dá)水平的影響。方法將研究對象(人肺腺癌A549細(xì)胞)分三組,分別是mi RNA-16轉(zhuǎn)染組,陰性序列轉(zhuǎn)染組,空白對照組。將50nmol/L的mi RNA-16 mimics和陰性對照序列用Lipofectamine 2000轉(zhuǎn)染進(jìn)人肺腺癌A549細(xì)胞中(空白對照組除了沒有轉(zhuǎn)染的RNA序列外,其他條件如所加的轉(zhuǎn)染混合液,實(shí)驗(yàn)對象等都與其他兩組一樣),通過q RT-PCR檢測轉(zhuǎn)染后A549細(xì)胞中mi RNA-16的表達(dá)水平,通過四甲基偶氮唑藍(lán)法(MTT法)檢測轉(zhuǎn)染mi RNA-16后不同時間點(diǎn)A549細(xì)胞的增殖抑制情況,通過ELSA法檢測轉(zhuǎn)染mi RNA-16并培養(yǎng)24小時后A549細(xì)胞上清液VEGF-A表達(dá)水平。結(jié)果q RT-PCR:轉(zhuǎn)染培養(yǎng)24小時后,測得轉(zhuǎn)染組mi RNA-16表達(dá)水平明顯高于空白對照組和陰性對照組(P0.05),其中轉(zhuǎn)染組表達(dá)水平是空白組的467.59±35.47倍,空白對照組和陰性對照組差異無統(tǒng)計(jì)學(xué)意義;MTT法:轉(zhuǎn)染并培養(yǎng)后,轉(zhuǎn)染組細(xì)胞增殖抑制率明顯高于空白對照組和陰性對照組(P0.05),在24小時,mi RNA-16轉(zhuǎn)染組、陰性序列轉(zhuǎn)染組和空白對照組的OD值分別是0.471±0.028、0.561±0.020和0.567±0.025,在48小時,三組的OD值分別是0.543±0.021、0.678±0.026和0.688±0.022,而在72小時,三組的OD值分別是0.623±0.016、0.895±0.019和0.907±0.018;陰性對照組和空白對照組比,差異無統(tǒng)計(jì)學(xué)意義;ELISA法:轉(zhuǎn)染并培養(yǎng)后,ELISA法測得轉(zhuǎn)染組細(xì)胞上清VEGF-A表達(dá)水平為55.56±6.04 pg/ml,較陰性對照組(152.07±6.41 pg/ml)和空白對照組(146.43±7.71 pg/ml)明顯降低(P0.05),而陰性對照組和空白對照組相比,差異無統(tǒng)計(jì)學(xué)意義。結(jié)論mi RNA-16能引起體外培養(yǎng)的人肺腺癌A549細(xì)胞增殖抑制,并且可能下調(diào)VEGF-A的表達(dá)。
[Abstract]:Objective to investigate the effects of transfection of mi RNA-16 on the proliferation of human lung adenocarcinoma A549 cells in vitro and the expression of VEGF-A in A549 cells. Methods Human lung adenocarcinoma A549 cells were divided into three groups: mi RNA-16 transfection group, negative sequence transfection group and blank control group. The 50 nmol / L mi RNA-16 mimics and the negative control sequence were transfected into human lung adenocarcinoma A549 cells by Lipofectamine 2000. The expression level of mi RNA-16 in A549 cells after transfection was detected by Q RT-PCR, and the proliferation inhibition of A549 cells was detected by MTT assay at different time points after transfection. The expression of VEGF-A in the supernatant of A549 cells was detected by ELSA method after transfection of mi RNA-16 and culture for 24 hours. Results Q RT-PCR: after 24 hours of transfection, the expression level of mi RNA-16 in the transfected group was significantly higher than that in the blank control group and the negative control group (P0.05), and the expression level in the transfected group was 467.59 鹵35.47 times higher than that in the blank group. There was no significant difference between the blank control group and the negative control group in MTT assay: after transfection and culture, the cell proliferation inhibition rate of the transfected group was significantly higher than that of the blank control group and the negative control group (P0.05). The OD values of the negative sequence transfection group and the blank control group were 0.471 鹵0.028, 0.561 鹵0.020 and 0.567 鹵0.025, respectively. At 48 hours, the OD values of the three groups were 0.543 鹵0.021, 0.678 鹵0.026 and 0.688 鹵0.022, respectively, while at 72 hours, the OD values of the three groups were 0.623 鹵0.0160.895 鹵0.019 and 0.907 鹵0.018, respectively. The expression of VEGF-A in the supernatant of the transfected group was 55.56 鹵6.04 PG / ml, which was significantly lower than that of the negative control group (152.07 鹵6.41 pg/ml) and the blank control group (146.43 鹵7.71 pg/ml) (P0.05). The difference was not statistically significant. Conclusion mi RNA-16 can inhibit the proliferation of human lung adenocarcinoma cell line A549 in vitro and may down-regulate the expression of VEGF-A.
【學(xué)位授予單位】：安徽醫(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

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉焱,楊重慶;老年人肺腺癌回腸轉(zhuǎn)移一例[J];中華老年醫(yī)學(xué)雜志;2005年05期

2 陳龍邦;;人肺腺癌多西他賽耐藥機(jī)制的研究[J];中國腫瘤生物治療雜志;2012年05期

3 杜鵑，孫培宗，李群英，張素珍，劉秋葵;老年人肺腺癌80例臨床分析[J];河南腫瘤學(xué)雜志;1994年04期

4 高振強(qiáng),高志萍,張濤,劉喜富;腫瘤特異表達(dá)的單純皰疹病毒胸苷激酶基因?qū)θ朔蜗侔┑膶?shí)驗(yàn)治療[J];中國科學(xué)C輯:生命科學(xué);1997年04期

5 潘世揚(yáng)，畢志剛，殷凱生，丁小健，，陳德才;維拉帕米影響絲裂霉素C抗人肺腺癌作用的體外研究[J];癌癥;1994年06期

6 胡義德,錢桂生,陳維中,肖桃元;人肺腺癌A_(549)多重抗藥細(xì)胞株生物學(xué)特性變化[J];第三軍醫(yī)大學(xué)學(xué)報;1997年05期

7 張峰,姜翠琴

本文編號：2093491