【摘要】：【目的】 檢測(cè)經(jīng)褪黑素處理后的膠質(zhì)瘤細(xì)胞株中microRNA-155(miR-155)的表達(dá)情況，探討褪黑素對(duì)膠質(zhì)瘤細(xì)胞增殖、轉(zhuǎn)移和侵襲能力的影響，并探究其抑癌的相關(guān)機(jī)制。 【方法】 第一部分：1、選用人腦膠質(zhì)瘤細(xì)胞株U87、U373、U251共三株，分別用不同濃度（100μM、1μM、1nM）的褪黑素處理。運(yùn)用qRT-PCR技術(shù)檢測(cè)三株膠質(zhì)瘤細(xì)胞中miR-155的表達(dá)量相對(duì)于對(duì)照組的變化。明確褪黑素和miR-155表達(dá)量的相關(guān)性，并確定引起miR-155表達(dá)量變化最大的褪黑素濃度。2、選定一株膠質(zhì)瘤細(xì)胞株，并選用引起miR-155表達(dá)量變化最大的濃度范圍的褪黑素處理細(xì)胞。通過(guò)CCK-8方法觀察膠質(zhì)瘤細(xì)胞的增殖情況。3、通過(guò)流式細(xì)胞儀技術(shù)檢測(cè)膠質(zhì)瘤細(xì)胞增殖、細(xì)胞周期及凋亡的變化。4、利用Transwell試驗(yàn)檢測(cè)褪黑素對(duì)腫瘤細(xì)胞遷移、侵襲的影響。 第二部分：1、選定一株膠質(zhì)瘤細(xì)胞，并選取引起miR-155表達(dá)量變化最大的濃度范圍的褪黑素處理細(xì)胞。分別應(yīng)用Western blot和qRT-PCR技術(shù)檢測(cè)細(xì)胞中MYB基因的mRNA及蛋白表達(dá)相對(duì)于對(duì)照組的變化，明確褪黑素與MYB基因的相關(guān)性。2、選同一株膠質(zhì)瘤細(xì)胞，將MYB基因干擾后用qRT-PCR技術(shù)檢測(cè)細(xì)胞中miR-155的表達(dá)量相對(duì)于對(duì)照組的變化，明確MYB和miR-155的相關(guān)性。 【結(jié)果】 第一部分：1、三株膠質(zhì)瘤細(xì)胞U87、U373、U251經(jīng)過(guò)不同濃度（100μM、1μM、1nM）褪黑素處理后，miR-155的表達(dá)相對(duì)于正常對(duì)照組均明顯下降(P0.05)，其中用1μM濃度褪黑素處理的細(xì)胞株中miR-155表達(dá)下降最明顯(P0.01)。2、經(jīng)過(guò)1μM濃度褪黑素處理的U87膠質(zhì)瘤細(xì)胞增殖較對(duì)照組明顯下降。3、經(jīng)過(guò)1μM濃度褪黑素處理的U87膠質(zhì)瘤細(xì)胞凋亡較對(duì)照組增高，而細(xì)胞周期沒(méi)有變化。4、經(jīng)過(guò)1μM濃度的褪黑素處理的U87膠質(zhì)瘤細(xì)胞遷移和侵襲能力較對(duì)照組明顯下降。 第二部分：1、經(jīng)過(guò)1μM濃度褪黑素處理的U87膠質(zhì)瘤細(xì)胞中MYB基因的mRNA和蛋白表達(dá)量均明顯低于對(duì)照組。2、在U87膠質(zhì)瘤細(xì)胞中干擾MYB基因后，miR-155的表達(dá)量下降。 【結(jié)論】 本研究結(jié)果表明褪黑素通過(guò)下調(diào)MYB基因從而抑制miR-155表達(dá)，1μM濃度褪黑素可以增加腫瘤細(xì)胞的凋亡使膠質(zhì)瘤細(xì)胞增殖降低，并且能抑制膠質(zhì)瘤細(xì)胞的遷移和侵襲能力，，進(jìn)而發(fā)揮抑癌作用。本實(shí)驗(yàn)闡明了褪黑素在腦膠質(zhì)瘤中抑癌作用的microRNA表觀調(diào)控機(jī)制，為揭示褪黑素的抗腦膠質(zhì)瘤作用機(jī)制奠定了理論基礎(chǔ)，為發(fā)現(xiàn)腦膠質(zhì)瘤的臨床治療靶點(diǎn)提供新思路。
[Abstract]:[objective] to detect the expression of microRNA-155 (miR-155) in glioma cells treated with melatonin, and to explore the effect of melatonin on the proliferation, metastasis and invasion of glioma cells. In the first part, three human glioma cell lines, U87U373U251, were treated with melatonin at different concentrations (100 渭 M, 1 渭 M, 1nM), respectively. The expression of miR-155 in three glioma cells was detected by qRT-PCR. To determine the correlation between the expression of miR-155 and melatonin, and to determine the concentration of melatonin. 2. To select a glioma cell line, and to select the melatonin treated cells in the concentration range that caused the most change of miR-155 expression. The proliferation of glioma cells was observed by CCK-8 method, the proliferation, cell cycle and apoptosis of glioma cells were detected by flow cytometry, and the effects of melatonin on the migration and invasion of glioma cells were detected by Transwell test. The second part: 1, a glioma cell line was selected, and melatonin treated cells were selected in the concentration range that caused the greatest change of miR-155 expression. The mRNA and protein expression of MYB gene were detected by Western blot and qRT-PCR. The correlation between melatonin and MYB gene was determined, and the same glioma cell line was selected. The expression of miR-155 was detected by qRT-PCR after MYB gene interference compared with the control group. The correlation between MYB and miR-155 was clarified. [results] in the first part, the expression of melatonin in U87U373U251 glioma cells was significantly lower than that in the control group (P0.05) after being treated with melatonin at different concentrations (100 渭 MU 1 渭 M) (P0.05). The expression of miR-155 in U87 glioma cells treated with 1 渭 M melatonin was significantly decreased (P0.01). The proliferation of U87 glioma cells treated with 1 渭 M melatonin was significantly lower than that of the control group. The apoptosis of U87 glioma cells treated with 1 渭 M melatonin was higher than that of the control. The cell cycle of U87 glioma cells treated with melatonin (1 渭 M) was not changed. The migration and invasion ability of U87 glioma cells treated with 1 渭 M melatonin was significantly lower than that of the control group. In the second part, the mRNA and protein expression of MYB gene in U87 glioma cells treated with melatonin at 1 渭 M concentration was significantly lower than that in control group, and the expression of miR-155 in U87 glioma cells was decreased after interfering with MYB gene in U87 glioma cells. [conclusion] this study suggests that melatonin can increase the apoptosis of tumor cells and decrease the proliferation of glioma cells by down-regulating the expression of miR-155 and inhibiting the expression of miR-155. And can inhibit the migration and invasion of glioma cells, and then play a role in cancer suppression. In this study, the mechanism of microRNA apparent regulation of melatonin in gliomas was elucidated, which laid a theoretical foundation for revealing the mechanism of melatonin's anti-glioma, and provided a new idea for the clinical treatment of glioma.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 方華鎣;許春芳;;褪黑素抗腫瘤作用及其機(jī)制的研究進(jìn)展[J];國(guó)際消化病雜志;2010年01期

2 徐學(xué)君,湯國(guó)太,游潮,張尚福,易章超;C-myb、PKC_a在腦膠質(zhì)瘤侵襲作用中的相關(guān)性研究[J];華西醫(yī)學(xué);2001年03期

3 胡世蓮;;褪黑素抗腫瘤作用及其機(jī)制的研究進(jìn)展[J];中國(guó)藥理學(xué)通報(bào);2008年04期

本文編號(hào)：2120191