本文關(guān)鍵詞： 骨腫瘤 骨肉瘤 miR-a SP JAK/STAT　出處：《臨床與實(shí)驗(yàn)病理學(xué)雜志》2016年10期 　論文類型：期刊論文

【摘要】：目的探討miR-135a通過(guò)靶向調(diào)節(jié)SP1對(duì)人骨肉瘤(osteosarcoma,OS)細(xì)胞增殖和凋亡的影響。方法收集人正常成骨組織和OS組織,培養(yǎng)正常成骨細(xì)胞(h FOB1.19)和OS細(xì)胞(MG-63),Real-time PCR法檢測(cè)miR-135a和SP1的表達(dá),Western blot法檢測(cè)SP1表達(dá)。轉(zhuǎn)染miR-135a mimics和inhibitor,MTT法和Brd U-ELISA法檢測(cè)OS細(xì)胞增殖變化,Western blot法檢測(cè)凋亡蛋白Bax、BCL-2和Caspase 3的表達(dá)。雙熒光素酶報(bào)告基因檢測(cè)miR-135a與SP1的靶定關(guān)系。Western blot法檢測(cè)miR-135a對(duì)OS細(xì)胞中SP1表達(dá)的影響。Western blot法檢測(cè)miR-135a對(duì)OS細(xì)胞中JAK2/STAT3表達(dá)的影響。結(jié)果 OS組織和細(xì)胞中miR-135a表達(dá)均顯著降低,SP1表達(dá)均顯著升高。轉(zhuǎn)染miR-135a mimics可降低OS細(xì)胞活性,減少Brd U陽(yáng)性標(biāo)記率。同時(shí),miR-135a mimics增加OS細(xì)胞Bax和Caspase 3的表達(dá),減少BCL-2的表達(dá)。miR-135a mimics降低熒光素酶報(bào)告基因的熒光強(qiáng)度,結(jié)合位點(diǎn)突變后熒光素酶活性升高。上調(diào)miR-135a顯著降低SP1的表達(dá)并降低JAK2和STAT3的磷酸化水平。miR-135a inhibitor作用均與mimics相反。結(jié)論 miR-135a可通過(guò)靶向調(diào)節(jié)SP1抑制人OS細(xì)胞增殖,誘導(dǎo)OS細(xì)胞凋亡,并下調(diào)JAK2/STAT3活化。
[Abstract]:Objective to investigate the effect of miR-135a on proliferation and apoptosis of human osteosarcoma osteosarcoma cells by targeting SP1. Methods normal human osteoblasts and OS tissues were collected. The expression of miR-135a and SP1 in cultured osteoblasts were detected by Western blot, and the proliferation of OS cells was detected by Brd U-ELISA and mimics transfection. The expression of Bax-BCL-2 and Caspase 3 were detected by Western blot method and the expression of Baxchon BCL-2 and Caspase 3 were detected by Western blot method. The expression of Baxchon BCL-2 and Caspase 3 were detected by Western blot method. The expression of Baxchon BCL-2 and Caspase 3 in cultured osteoblasts were detected by Western blot. Double luciferase reporter gene was used to detect the target relationship between miR-135a and SP1. Western blot method was used to detect the effect of miR-135a on SP1 expression in OS cells. Western blot assay was used to detect the effect of miR-135a on JAK2/STAT3 expression in OS cells. Transfection of miR-135a mimics decreased the activity of OS cells. At the same time, miR-135a mimics increased the expression of Bax and Caspase 3 in OS cells, and decreased the expression of BCL-2. MiR-135a mimics decreased the fluorescence intensity of luciferase reporter gene. After binding site mutation, luciferase activity increased. Upregulation of miR-135a significantly decreased the expression of SP1 and decreased the phosphorylation level of JAK2 and STAT3. MIR-135a inhibitor had the opposite effect on mimics. Conclusion miR-135a can inhibit the proliferation of human OS cells by targeting SP1. The apoptosis of OS cells was induced and the activation of JAK2/STAT3 was down-regulated.
【作者單位】： 河南省中醫(yī)院骨病一科;商丘醫(yī)學(xué)高等�？茖W(xué)校臨床系中醫(yī)教研室;
【分類號(hào)】：R738.1
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本文編號(hào)：1551941