本文選題：MiR- + 骨肉瘤　； 參考：《中華腫瘤防治雜志》2017年16期

【摘要】：目的骨肉瘤具有局部高度侵襲能力以及快速轉(zhuǎn)移潛能,因而致死率較高。研究指出,miR-300的表達(dá)異常與多種腫瘤的發(fā)病相關(guān)。miR-300對(duì)骨肉瘤細(xì)胞是否存在調(diào)控作用卻屬未知。本研究探討miR-300對(duì)骨肉瘤細(xì)胞Saos-2增殖與凋亡調(diào)控的作用。方法將miR-300轉(zhuǎn)染進(jìn)骨肉瘤細(xì)胞Saos-2中,實(shí)現(xiàn)miR-300在Saos-2細(xì)胞內(nèi)的高表達(dá)。通過(guò)熒光定量PCR測(cè)定miR-300在細(xì)胞內(nèi)的表達(dá)水平;分別采用MTT活細(xì)胞測(cè)試,細(xì)胞周期實(shí)驗(yàn)和克隆形成實(shí)驗(yàn)檢測(cè)骨髓瘤細(xì)胞的增殖情況;通過(guò)蛋白質(zhì)印跡法測(cè)定Bcl-2、Bax和Bak,裂解型Caspase-3蛋白水平來(lái)鑒定細(xì)胞凋亡。結(jié)果轉(zhuǎn)染miR-300的Saos-2細(xì)胞,其miR-300表達(dá)量為對(duì)照組的(10.23±1.04)倍,t=3.613 8,P=0.000 6。MTT實(shí)驗(yàn)結(jié)果顯示,在轉(zhuǎn)染后24和48h,miR-300組的Saos-2相對(duì)活細(xì)胞百分比相對(duì)對(duì)照組分別為[(174.35±28.46)%,t=2.219,P=0.03]和[(225.73±24.62)%,t=2.738,P=0.008]。miR-300組Saos-2細(xì)胞處在G1期的百分比為39.42%,低于對(duì)照組的47.58%,t=2.366,P=0.021;而處在G2期的細(xì)胞百分比為19.12%,顯著高于對(duì)照組的10.55%,t=2.987,P=0.004。miR-300組Saos-2細(xì)胞克隆形成率為(52.53±30.21)%,顯著高于對(duì)照組的(24.67±2.05)%,t=2.711,P=0.008 6。miR-300組Saos-2細(xì)胞中Bax、Bak和裂解型Caspase-3表達(dá)水平分別為對(duì)照組的(0.25±0.02)倍(t=2.785,P=0.007)、(0.31±0.03)倍(t=3.223,P=0.002)和(0.36±0.03)倍(t=3.006,P=0.003 8),差異有統(tǒng)計(jì)學(xué)意義。Bcl-2蛋白水平為對(duì)照組的(6.32±0.57)倍,t=3.218,P=0.002。轉(zhuǎn)染miRZip lent-shMiR-300的Saos-2細(xì)胞,其miR-300數(shù)量為對(duì)照組的(0.28±0.03)倍,t=3.531,P=0.000 78。轉(zhuǎn)染24h后,miR-300組Saos-2相對(duì)活細(xì)胞數(shù)為對(duì)照組的(64.46±5.32)%,t=2.324,P=0.024;48h后為對(duì)照組的(48.14±4.38)%,t=3.009,P=0.004。miR-300組Saos-2細(xì)胞處在G1期的百分比為55.71%,高于對(duì)照組的46.78%,t=2.108,P=0.039;處在G2期的細(xì)胞百分比為2.81%,低于對(duì)照組的11.46%,t=3.224,P=0.002。miR-300組的Saos-2細(xì)胞的克隆形成率為(17.63±3.89)%,顯著低于對(duì)照組的(26.84±2.94)%,t=2.989,P=0.004。miR-300組Saos-2細(xì)胞中Bax表達(dá)水平為對(duì)照組的(6.25±4.23)倍,t=3.138,P=0.002 6;Bak表達(dá)水平為對(duì)照組的(6.03±4.27)倍,t=3.395,P=0.001 2;裂解型Capase-3表達(dá)水平為對(duì)照組的(5.35±0.37)倍,t=3.017,P=0.003 7;而B(niǎo)cl-2蛋白水平為對(duì)照組的(0.36±0.02)倍,t=2.769,P=0.007 4。結(jié)論 miR-300在骨肉瘤細(xì)胞的增殖與凋亡調(diào)控中具有重要作用,抑制miR-300表達(dá)可以一定程度上減少骨髓瘤細(xì)胞的增殖,促進(jìn)其凋亡,為骨肉瘤的病理機(jī)制研究提供了新研究方向。
[Abstract]:Objective Osteosarcoma has high local invasion and rapid metastasis potential, so the fatality rate is high.It is suggested that the abnormal expression of miR-300 is related to the pathogenesis of many kinds of tumors. Whether miR-300 can regulate osteosarcoma cells is unknown.To investigate the effect of miR-300 on proliferation and apoptosis of osteosarcoma cell line Saos-2.Methods miR-300 was transfected into Saos-2 of osteosarcoma cells, and the high expression of miR-300 in Saos-2 cells was realized.The expression of miR-300 was measured by fluorescence quantitative PCR, and the proliferation of myeloma cells was detected by MTT living cell test, cell cycle assay and clone formation assay.Apoptotic cells were identified by Western blotting assay of Bcl-2P Bax and Bak-Bax and cleavage Caspase-3 protein levels.Results the expression of miR-300 in Saos-2 cells transfected with miR-300 was 10.23 鹵1.04 times as high as that in the control group. The results of the experiment showed that the expression of miR-300 was 3.6138Pu 0.000 6.MTT.鐨，

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