本文關(guān)鍵詞：miR-483-3P下調(diào)RIOK3蛋白表達(dá)并促進(jìn)神經(jīng)母細(xì)胞瘤的增殖侵襲和遷移　出處：《青島大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 神經(jīng)母細(xì)胞瘤 mi R-483-3P 細(xì)胞增殖 侵襲 靶基因

【摘要】：目的通過實(shí)驗(yàn)研究micro RNA-483-3P(mi R-483-3P)對(duì)神經(jīng)母細(xì)胞瘤細(xì)胞的生物學(xué)行為的影響,諸如增殖能力、侵襲能力及遷移能力。利用生物信息學(xué)軟件預(yù)測(cè)mi R-483-3P的靶基因并加以驗(yàn)證,以及探索mi R-483-3P對(duì)所預(yù)測(cè)的靶基因的影響。方法mi RNA微陣列芯片(Micro Array)的結(jié)果發(fā)現(xiàn),與原發(fā)瘤相比,mi R-483-3P在神經(jīng)母細(xì)胞轉(zhuǎn)移瘤中表達(dá)水平明顯上調(diào),差異倍數(shù)顯著,并居于所檢測(cè)出的差異表達(dá)的mi RNA的第一位。利用陽(yáng)離子脂質(zhì)體Lipofectamine TM2000將化學(xué)合成的mi R-483-3P inhibitor、mi R-483-3P inhibitor的陰性對(duì)照序列(negative control)分別轉(zhuǎn)染入人神經(jīng)母細(xì)胞瘤SH-SY-5Y細(xì)胞株中,實(shí)時(shí)熒光定量聚合酶鏈?zhǔn)椒磻?yīng)(RT-q PCR)技術(shù)檢測(cè)各組神經(jīng)母細(xì)胞瘤經(jīng)處理后mi R-483-3P的表達(dá)水平,細(xì)胞計(jì)數(shù)試劑盒(CCK-8)法檢測(cè)各組神經(jīng)母細(xì)胞瘤細(xì)胞的增殖情況,Transwell小室實(shí)驗(yàn)檢測(cè)各組神經(jīng)母細(xì)胞瘤細(xì)胞的體外侵襲以及遷移能力。利用多個(gè)靶基因預(yù)測(cè)數(shù)據(jù)庫(kù),綜合預(yù)測(cè)mi R-483-3P的靶基因,熒光素酶報(bào)告基因檢測(cè)實(shí)驗(yàn)、Western blot實(shí)驗(yàn)加以驗(yàn)證。結(jié)果實(shí)驗(yàn)所取的9對(duì)神經(jīng)母細(xì)胞瘤組織與癌旁正常組織,RT-QPCR結(jié)果顯示,mi R-483-3P在神經(jīng)母細(xì)胞瘤腫瘤組織中表達(dá)水平明顯高于癌旁正常組織(P0.01);與轉(zhuǎn)染了mi R-483-3P inhibitor的陰性對(duì)照組相比,癌細(xì)胞轉(zhuǎn)染mi R-483-3P inhibitor后,mi R-483-3P被抑制表達(dá)水平顯著下調(diào)(P0.01),細(xì)胞的增殖情況和體外遷移能力均降低(P0.05)。經(jīng)多個(gè)靶基因生物信息學(xué)軟件分析,預(yù)測(cè)出RIOK3是mi R-483-3P的靶基因之一,當(dāng)神經(jīng)母細(xì)胞瘤細(xì)胞轉(zhuǎn)染了mi R-483-3P inhibitor后熒光酶的活性明顯升高(P0.01)。與mi R-483-3P inhibitor陰性對(duì)照組相比,當(dāng)細(xì)胞轉(zhuǎn)染了mi R-483-3P inhibitor后,RIOK3蛋白的表達(dá)水平明顯升高(P0.01)。結(jié)論RIOK3是miR-483-3P的靶基因之一,miR-483-3P能下調(diào)RIOK3蛋白的表達(dá),mi R-483-3P可顯著促進(jìn)神經(jīng)母細(xì)胞瘤細(xì)胞的增殖及體外遷移能力。
[Abstract]:Objective to investigate the effects of micro RNA-483-3P(mi R-483-3P on the biological behavior of neuroblastoma cells, such as proliferation. Invasive ability and migration ability. The target gene of mi R-483-3P was predicted and verified by bioinformatics software. And to explore the effect of mi R-483-3P on the predicted target gene. Methods the results of mi RNA microarray microarray microarray showed that compared with the primary tumor. The expression of mi R-483-3P was up-regulated in neuroblastoma, and the difference was significant. The chemically synthesized mi R-483-3P was chemically synthesized using cationic liposome Lipofectamine TM2000. Inhibitor. Negative control sequence of mi R-483-3P inhibitor. The cells were transfected into human neuroblastoma SH-SY-5Y cell lines. Real-time quantitative polymerase chain reaction (RT-q PCR) technique was used to detect the expression of mi R-483-3P in neuroblastoma after treatment. The proliferation of neuroblastoma cells in each group was detected by CCK-8 assay. The invasiveness and migration ability of neuroblastoma cells in each group were detected by Transwell chamber assay. The target genes of mi R-483-3P were comprehensively predicted by using multiple target gene prediction databases. Results 9 pairs of neuroblastoma tissues and adjacent normal tissues were detected by RT-QPCR. The expression of miR-483-3P in neuroblastoma was significantly higher than that in adjacent normal tissue (P 0.01). Compared with the negative control group transfected with mi R-483-3P inhibitor, the cancer cells were transfected with mi R-483-3P inhibitor. Mi R-483-3P was significantly down-regulated (P0.01). The cell proliferation and migration ability in vitro were decreased (P 0.05). It was predicted that RIOK3 was one of the target genes of mi R-483-3P by multiple target gene bioinformatics software analysis. The activity of fluorescence enzyme increased significantly after transfection of mi R-483-3P inhibitor into neuroblastoma cells (P 0.01). Compared with mi R-483-3P inhibitor negative control group. When the cells were transfected with mi R-483-3P inhibitor. Conclusion RIOK3 is one of the target genes of miR-483-3P. MiR-483-3P can down-regulate the expression of RIOK3 protein. Mi R-483-3P can significantly promote the proliferation and migration of neuroblastoma cells in vitro.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R739.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 周顯軍;陳鑫;沈峰;董劏;朱永潔;段于河;耿耿;崔楷悅;李曉;;miR-142-3p調(diào)控高遷移率族蛋白1及對(duì)神經(jīng)母細(xì)胞瘤順鉑化療敏感性的影響[J];中華小兒外科雜志;2015年06期

2 劉媛;蔣菁蕊;吳少亙;曹望森;于成功;崔恒宓;;miR-483與結(jié)直腸癌關(guān)系的研究[J];胃腸病學(xué);2014年03期

3 林穎;李璞;單敬軒;陳曉靜;施慧莉;霍克克;;RIOK3促進(jìn)了caspase-10對(duì)PAK2的酶解激活[J];中國(guó)生物工程雜志;2012年08期

4 宋華;董劏;呂振華;侯琳;孫立榮;王偉;鹿洪亭;郝希偉;;KAI1/CD82在神經(jīng)母細(xì)胞瘤轉(zhuǎn)移中的表達(dá)及意義[J];中華小兒外科雜志;2007年08期

5 張桓瑜;鹿洪亭;董劏;楊傳民;郝希偉;蘇南;;微小RNA-7對(duì)人神經(jīng)母細(xì)胞瘤侵襲和轉(zhuǎn)移能力的影響[J];中華實(shí)驗(yàn)外科雜志;2013年06期

6 鹿洪亭;李富江;康曉寧;丁玉虎;陳鑫;張桓瑜;蘇南;董劏;;miRNAs的表達(dá)在神經(jīng)母細(xì)胞瘤轉(zhuǎn)移過程中的作用及意義[J];中華小兒外科雜志;2013年06期

7 鹿洪亭;董劏;陳鑫;張虹;郝希偉;韓蘆蘆;;miR-338-3p對(duì)神經(jīng)母細(xì)胞瘤增殖和侵襲的抑制作用[J];中華小兒外科雜志;2013年07期

，

本文編號(hào)：1359018