本文選題：甲狀腺癌 + miRNA　； 參考：《中國老年學(xué)雜志》2017年03期

【摘要】：目的探討miR-124對甲狀腺癌細胞的作用機制。方法通過RT-PCR檢測人甲狀腺癌細胞K1、BCPAP、TPC-1和人甲狀腺細胞Nthy-ori 3-1中miR-124的表達水平。MTT法檢測甲狀腺癌細胞K1轉(zhuǎn)染miR-124 mimics、mimics control后的細胞存活率。根據(jù)靶基因預(yù)測軟件The miRBase、Target Scan、Pic Tar預(yù)測miR-124的靶基因IQGAP1,構(gòu)建wt-IQGAP1和mut-IQGAP1,采用雙熒光素酶活性檢測鑒定靶基因的正確性。Western印跡檢測轉(zhuǎn)染miR-124 mimics、mimics control后甲狀腺癌細胞K1中IQGAP1的表達情況。甲狀腺癌細胞K1中轉(zhuǎn)染si IQGAP1和si IQGAP1 control,用MTT檢測細胞增殖能力,Western印跡檢測細胞中IQGAP1蛋白表達含量。結(jié)果甲狀腺癌細胞K1、BCPAP、TPC-1中的miR-124的相對表達量與甲狀腺細胞相比差異顯著(P0.05),三種甲狀腺癌細胞中K1細胞的表達量最低。轉(zhuǎn)染后12 h內(nèi),空白組、miR-124 mimics組和mimics control組的甲狀腺癌K1細胞的存活數(shù)量差異不顯著(P0.05),從轉(zhuǎn)染后12 h開始,miR-124 mimics組與對照組、mimics control組相比,甲狀腺癌細胞生長抑制明顯。預(yù)測miR-124的靶基因可能為IQGAP1,轉(zhuǎn)染miR-124 mimics野生型IQGAP1中熒光素酶活性比突變型IQGAP1中熒光素酶活性顯著降低,差異顯著(P0.05)。mimics control和wt-IQGAP1和mut-IQGAP1共轉(zhuǎn)染組比較無明顯差異(P0.05)。轉(zhuǎn)染miR-124 mimics組甲狀腺癌K1細胞IQGAP1蛋白明顯下降,miR-124負調(diào)控IQGAP1的表達。抑制甲狀腺癌K1細胞中IQGAP1基因,細胞增殖能力減弱,細胞中IQGAP1蛋白表達減弱。結(jié)論 miR-124在甲狀腺癌細胞中表達下調(diào),miR-124可以抑制甲狀腺癌細胞的增殖分化,其通過負調(diào)控靶基因IQGAP1抑制癌細胞增殖分化。
[Abstract]:Objective to investigate the mechanism of miR-124 acting on thyroid cancer cells. Methods the expression level of miR-124 in human thyroid cancer cell K1 / BCPAPP / TPC-1 and human thyroid cell Nthy-ori 3-1 was detected by RT-PCR. The survival rate of K1 transfected with miR-124 mimicsmimics control was detected by MTT assay. The target gene IQGAP1 of miR-124 was predicted by the target gene prediction software, the miRBaseScanTar. Wt-IQGAP1 and mut-IQGAP1 were constructed. The expression of IQGAP1 in thyroid cancer cell K1 after transfection of miR-124 mimicsmimics control was detected by double luciferase activity assay. Si IQGAP1 and si IQGAP1 control1 were transfected into thyroid cancer cell K1 and the expression of IQGAP1 protein was detected by MTT assay and Western blotting. Results the relative expression of miR-124 in thyroid cancer cell K1 BCPAPAPP TPC-1 was significantly different from that in thyroid cell (P0.05), and the expression of K1 in three kinds of thyroid cancer cells was the lowest. Within 12 hours after transfection, there was no significant difference in the survival of K1 cells between the blank group and the mimics control group (P0.05). From 12 h after transfection, the growth inhibition of thyroid cancer cells in the miR-124 mimics group was significantly higher than that in the control group (mimics control group). It was predicted that the target gene of miR-124 might be IQGAP1. The luciferase activity of transfected miR-124 mimics wild-type IQGAP1 was significantly lower than that of mutant IQGAP1 (P0.05). There was no significant difference between mimics control and wt-IQGAP1 and mut-IQGAP1 co-transfection groups (P0.05). The expression of IQGAP1 in K1 cells of thyroid carcinoma was significantly decreased by miR-124 mimics transfection, and the expression of IQGAP1 was negatively regulated by miR-124. Inhibition of IQGAP1 gene in K1 cells decreased cell proliferation and IQGAP1 protein expression. Conclusion the down-regulated expression of miR-124 in thyroid cancer cells can inhibit the proliferation and differentiation of thyroid cancer cells, and it can inhibit the proliferation and differentiation of thyroid cancer cells through negative regulation target gene IQGAP1.
【作者單位】： 天津市海河醫(yī)院內(nèi)科;天津市醫(yī)科大學(xué)總醫(yī)院內(nèi)分泌科;
【分類號】：R736.1

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