抑制硬化蛋白基因表达有利于诱导成骨细胞的分化:体外乳腺癌骨转移模型

发布时间：2018-05-04 10:32

本文选题：硬化蛋白 + 乳腺癌　；参考：《南方医科大学学报》2017年08期

【摘要】：目的探讨在体外乳腺癌骨转移模型中抑制硬化蛋白基因的表达后对成骨细胞分化成熟的影响。方法以干扰硬化蛋白基因表达的si RNA腺病毒感染乳腺癌细胞株MDA-MB-231,RT-PCR检测确认硬化蛋白基因表达的改变。分别以正常培养的MDA-MB-231上清液,感染空病毒的MDA-MB-231上清液,感染SiSOST病毒的MDA-MB-231上清液与MG63或C3H10共培养,并以成骨诱导培养基培养MG63及C3H10为阳性对照,普通培养基培养MG63及C3H10为阴性对照。定量PCR检测成骨细胞分化相关指标的改变,染色检测碱性磷酸酶(ALP)表达的改变。结果感染Ad-si SOST病毒的MDA-MB-231细胞中硬化蛋白表达明显降低。定量PCR结果显示:与单独培养的MG63相比,加成骨培养基后其骨钙素、骨桥蛋白、护骨因子和整合素结合涎蛋白的表达均上升(P0.01)。在此基础上,与231细胞上清液共培养后其表达降低(P0.01),差异具有统计学意义;MG63+231-RFP组结果一致,感染SiSOST病毒后,分化指标重新上升(P0.01),差异具有统计学意义。C3H10细胞实验结果趋势一致。MG63细胞与阴性对照组相比,分化诱导培养基可促进ALP的分泌(P0.01),但与231细胞共培养后,ALP分泌减少(P0.01),感染Si-SOST后,可部分扭转ALP分泌的减少(P0.01),ALP染色得到同样的结果。C3H10细胞结果与MG63细胞趋势一致。结论在体外乳腺癌骨转移模型中,成骨细胞分化受阻;干扰硬化蛋白基因的表达有利于逆转成骨细胞的分化受阻。
[Abstract]:Objective to investigate the effect of inhibition of sclerosin gene expression on differentiation and maturation of osteoblasts in bone metastases of breast cancer in vitro. Methods the expression of sclerosin gene was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) in breast cancer cell line MDA-MB-231infected with si RNA adenovirus which interfered with sclerosing protein gene expression. MDA-MB-231 supernatant of normal culture, MDA-MB-231 supernatant infected with empty virus, MDA-MB-231 supernatant infected with SiSOST virus co-cultured with MG63 or C3H10, and MG63 and C3H10 were cultured in osteogenic medium as positive control. MG63 and C3H10 in normal culture medium were negative control. Quantitative PCR was used to detect the changes of osteoblast differentiation and alkaline phosphatase (ALP) expression was detected by staining. Results the expression of sclerosin in MDA-MB-231 cells infected with Ad-si SOST virus was significantly decreased. The results of quantitative PCR showed that the expression of osteocalcin, osteopontin, osteopontin, osteoprotegerin and integrin-binding sialoprotein increased after addition of bone medium compared with MG63 cultured alone. On this basis, after co-culture with the supernatant of 231 cells, the expression of P0.01was decreased, and the difference was consistent with that of MG63 231-RFP group. After infection of MG63 231-RFP, the expression of MG63 231-RFP was similar to that of MG63 virus. Compared with the negative control group, the differentiation induction medium could promote the secretion of ALP, but after co-culture with 231 cells, the secretion of P0.01C was decreased, and after infection with Si-SOST, the differentiation and induction medium could decrease the secretion of P0.01C _ 3H _ (10) cells, and the results showed that the differentiation of MG63 cells was similar to that of the negative control group (P _ (0.01), but the differentiation induction medium could promote the secretion of ALP. The same result could be obtained by partially reversing the decrease of ALP secretion. The result of C3H10 cells was consistent with that of MG63 cells. Conclusion in the model of bone metastasis of breast cancer in vitro, the differentiation of osteoblasts is blocked, and the expression of interfering sclerosing protein gene is beneficial to reverse the obstruction of osteoblast differentiation.
【作者单位】：重庆医科大学病理生理学教研室;重庆医科大学分子医学与肿瘤研究中心;重庆医科大学附属第二医院三腺外科;
【基金】：重庆市科技计划项目(cstc2013jcyj A1009)
【分类号】：R737.9

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