本文選題：mTORC信號 + 成骨分化��； 參考：《中國免疫學(xué)雜志》2016年11期

【摘要】：目的:研究mTORC1信號對前成骨細胞MC3T3-E1向成骨細胞分化成熟的調(diào)控作用。方法:通過向MC3T3-E1轉(zhuǎn)染pcDNA3.1-Raptor,對mTORC1信號相關(guān)蛋白Raptor進行過表達。向MC3T3-E1轉(zhuǎn)染Raptor siRNA,對mTORC1信號蛋白Raptor進行基因沉默。通過Real-time PCR方法測定Raptor的基因表達,通過蛋白免疫印跡法測定Raptor蛋白水平,并通過茜素紅染色檢測成骨礦化情況,以測定成骨分化程度。通過Real-time PCR檢測成骨分化指標的基因表達。結(jié)果:與對照組相比,Raptor過表達組的Raptor mRNA和蛋白水平明顯增加;茜素紅染色結(jié)果顯示Raptor過表達組染色更深,說明成骨礦化程度更高;熒光定量PCR結(jié)果顯示,Raptor過表達組的成骨分化標記基因以及成骨轉(zhuǎn)錄因子的表達量均高于對照組。與對照組相比,Raptor siRNA組的Raptor mRNA和蛋白水平明顯降低;茜素紅染色結(jié)果顯示Raptor siRNA組染色更淺,說明成骨礦化程度更低;熒光定量PCR結(jié)果顯示,Raptor siRNA組的成骨分化標記基因以及成骨轉(zhuǎn)錄因子的表達量均低于對照組。結(jié)論:mTORC1信號促進前成骨細胞MC3T3-E1向成骨細胞分化成熟。
[Abstract]:Aim: to study the effect of mTORC1 signal on the differentiation and maturation of preosteoblast MC3T3-E1 into osteoblasts. Methods: mTORC1 signal-associated protein (Raptor) was overexpressed by transfection of pcDNA3.1-Raptor. into MC3T3-E1. Raptor siRNAs were transfected into MC3T3-E1 and the mTORC1 signal protein Raptor was silenced. The gene expression of Raptor was detected by Real-time PCR method, the level of Raptor protein was detected by Western blot, and the osteogenic mineralization was detected by alizarin red staining to determine the degree of osteogenic differentiation. The gene expression of osteogenic differentiation index was detected by Real-time PCR. Results: compared with the control group, the levels of Raptor mRNA and protein were significantly increased in the Raptor overexpression group, and alizarin red staining showed that the Raptor overexpression group was more stained, indicating that the degree of osteogenesis mineralization was higher. The results of fluorescence quantitative PCR showed that the expression of osteogenic differentiation marker gene and osteogenic transcription factor in Raptor overexpression group was higher than that in control group. Compared with the control group, the levels of Raptor mRNA and protein in Raptor siRNA group were significantly lower, and alizarin red staining showed that Raptor siRNA group was lighter, indicating that the degree of osteogenesis mineralization was lower. The results of fluorescence quantitative PCR showed that the expression of osteogenic differentiation marker gene and osteogenic transcription factor in Raptor siRNA group was lower than that in control group. Conclusion the MC3T3-E1 of preosteoblast can be differentiated and matured by the signal of 1: mTORC1.
【作者單位】： 河南省新鄉(xiāng)醫(yī)學(xué)院醫(yī)學(xué)檢驗學(xué)院和分子診斷與醫(yī)學(xué)檢驗技術(shù)河南省協(xié)同創(chuàng)新中心;河南省新鄉(xiāng)醫(yī)學(xué)院;河南省新鄉(xiāng)醫(yī)學(xué)院第三附屬醫(yī)院;
【基金】：國家自然科學(xué)基金項目(No.31301135)
【分類號】：R329.2

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