【摘要】：目的檢測(cè)缺氧環(huán)境下人膠質(zhì)瘤A172細(xì)胞系中HIF-1、BNIP3的表達(dá)及兩者之間的關(guān)系，并探討其對(duì)膠質(zhì)瘤細(xì)胞的抑制增殖和促凋亡作用。 方法1.培養(yǎng)人膠質(zhì)母細(xì)胞瘤A172細(xì)胞系并制作缺氧箱模型，并將其隨機(jī)分為A組(空白對(duì)照組)、B組（缺氧組）、C組（缺氧+HIF-1激動(dòng)劑）、D組（缺氧+HIF-1激動(dòng)劑+BNIP-3抑制劑）共4組，相差顯微鏡觀察各組細(xì)胞在24h、48h、72h時(shí)間點(diǎn)的細(xì)胞形態(tài)學(xué)變化，MTT法檢測(cè)各組細(xì)胞增殖抑制情況。2. Tunel熒光一步法檢測(cè)各組A172細(xì)胞凋亡和增殖。3.運(yùn)用細(xì)胞免疫組化方法檢測(cè)各組細(xì)胞中HIF-1與BNIP-3的蛋白表達(dá)情況。 結(jié)果1.相差顯微鏡觀察：與A組比較，B、C、D三組細(xì)胞存活減少，增殖受到抑制，以B、C兩組為著。MTT法：B、C、D組細(xì)胞增殖受到抑制，以B、C兩組為著。2.Tunel熒光法檢測(cè)：與A組相比，B、C兩組細(xì)胞存活增殖差異有顯著統(tǒng)計(jì)學(xué)意義（P0.05）；與B組比較，C組細(xì)胞存活增殖差異有統(tǒng)計(jì)學(xué)意義（P0.05）；與C組比較，D組細(xì)胞存活增殖差異有統(tǒng)計(jì)學(xué)意義(P0.05)；3.免疫組化檢測(cè)：(1)蛋白HIF-1表達(dá)：與A組比較，B、C、D三組HIF-1的表達(dá)明顯增高，差異具有統(tǒng)計(jì)學(xué)意義（P0.05）；與B組比較，C、D兩組中HIF-1的表達(dá)增高，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。(2)蛋白BNIP-3表達(dá)：D組BNIP-3表達(dá)與B、C兩組比較明顯減少，差異有統(tǒng)計(jì)學(xué)意義（P0.05），A、B、C、D四組各時(shí)間點(diǎn)BNIP-3表達(dá)與HIF-1表達(dá)呈正相關(guān)，具有統(tǒng)計(jì)學(xué)意義。 結(jié)論1.人膠質(zhì)瘤A172細(xì)胞系在缺氧環(huán)境下增殖可受到抑制，細(xì)胞發(fā)生凋亡；2.HIF-1在缺氧環(huán)境下表達(dá)增高，，并可誘導(dǎo)BNIP-3的生成，兩者呈正相關(guān)關(guān)系；3.HIF-1可誘導(dǎo)A172細(xì)胞發(fā)生凋亡，可能的機(jī)制可能與其誘導(dǎo)BNIP-3的生成有關(guān)。
[Abstract]:Aim to investigate the expression of HIF-1,BNIP3 in human glioma cell line A172 under hypoxia and its relationship, and to explore its inhibitory effect on proliferation and apoptosis of human glioma cell line A172. Method 1. Human glioblastoma cell line A172 was cultured and hypoxia box model was made. They were randomly divided into group A (blank control group), B) (hypoxia group), C group (hypoxia HIF-1 agonist). Group D (hypoxia HIF-1 agonist BNIP-3 inhibitor) was divided into 4 groups. Phase contrast microscope was used to observe the morphological changes of the cells at 24 h, 48 h, 72 h, and MTT assay was used to detect the inhibition of cell proliferation. 2. Detection of apoptosis and proliferation of A172 cells by Tunel fluorescence one-step method. 3. The protein expression of HIF-1 and BNIP-3 in the cells of each group was detected by immunocytochemistry. Outcome 1. Phase contrast microscope observation: compared with group A, three groups B, C, D had a decrease in survival and proliferation, especially in B and C groups. MTT method: B, C, D group cell proliferation was inhibited, B, C, D groups were inhibited, B, C, D group cell proliferation was inhibited, B, C, D group were inhibited by B, C, D. 2.Tunel fluorescence assay: compared with group A, there was significant difference in cell survival and proliferation between group B and group C (P0.05). Compared with group B, group C had significant difference in cell survival and proliferation (P0.05); group D had significant difference in cell survival and proliferation compared with group C (P0.05); Immunohistochemistry: (1) the expression of protein HIF-1: compared with group A, the expression of HIF-1 in group B, C and D was significantly higher than that in group A (P0.05). Compared with group B, the expression of HIF-1 in group C and D was significantly higher than that in group B (P0.05). (2). The expression of BNIP-3 in group D was significantly lower than that in group B and C (P < 0.05). The difference was statistically significant (P0.05). There was a positive correlation between BNIP-3 expression and HIF-1 expression at each time point in A, B, C, D groups. Conclusion 1. The proliferation and apoptosis of human glioma cell line A172 were inhibited in hypoxia environment, and the expression of 2.HIF-1 was increased in hypoxia environment, and the production of BNIP-3 was induced, and there was a positive correlation between them. 3.HIF-1 can induce apoptosis of A172 cells, and the possible mechanism may be related to the induction of BNIP-3 production.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.41

【參考文獻(xiàn)】

相關(guān)期刊論文 前6條

1 許文偉;付俊武;闞文宏;;缺氧誘導(dǎo)因子-1α對(duì)腫瘤調(diào)控機(jī)制的研究進(jìn)展[J];廣東醫(yī)學(xué);2011年22期

2 李勇堅(jiān);祖旭宇;張桂英;肖嶸;文海泉;;低氧誘導(dǎo)因子-1α siRNA對(duì)HaCaT細(xì)胞誘導(dǎo)型一氧化氮合酶表達(dá)的影響[J];中南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2011年10期

3 劉林;薛文;錢耀文;;缺氧對(duì)乳鼠脊髓神經(jīng)元低氧誘導(dǎo)因子-1α的影響[J];中華神經(jīng)外科疾病研究雜志;2011年05期

4 呂寶軍;侯冰宗;茍新敏;王曉鴻;;乳腺浸潤(rùn)性導(dǎo)管癌中微淋巴管密度與缺氧誘導(dǎo)因子-1α的表達(dá)及其相關(guān)性[J];中國(guó)臨床研究;2011年09期

5 楊峰;湯欣;;低氧誘導(dǎo)因子-1α誘導(dǎo)鼠胚成骨細(xì)胞血管內(nèi)皮生長(zhǎng)因子(VEGF)表達(dá)的實(shí)驗(yàn)研究[J];中國(guó)傷殘醫(yī)學(xué);2011年08期

6 李劍敏,吳秀玲,朱啟建,王群姬;腦星形細(xì)胞腫瘤組織中內(nèi)皮素-1的表達(dá)及定量分析[J];癌癥;2002年10期

本文編號(hào)：2472201