论文部分内容阅读
目的通过共培养体系探讨成骨前体细胞MC3T3-E1对MDA-MB-231乳腺癌细胞表达骨拟态特性及增殖的影响。方法实验组将MC3T3-E1及MDA-MB-231细胞采用transwell小室共培养,上室接种MC3T3-E1成骨细胞,下室接种相同数量的MDA-MB-231细胞。对照组上、下室均采用相同数量及密度的MDA-MB-231细胞。实时荧光定量PCR、Western blot法检测实验组及对照组骨钙蛋白(osteocalcin,OCN)、骨桥蛋白(osteopontin,OPN)、骨保护素(osteoprotegerin,OPG)的基因及蛋白的表达差异,采用CCK-8法检测各组增殖的差异,结晶紫染色法检测细胞集落形成差异。结果与对照组相比,实验组OCN、OPN、OPG的基因及蛋白表达均有明显的上调(P<0.05),MDA-MB-231细胞的增殖能力明显增高(P<0.01);实验组MDA-MB-231细胞集落形成数量及大小均优于对照组。结论短期共培养后MC3T3-E1成骨细胞可促进乳腺癌MDA-MB-231细胞表达骨拟态特性,并可促进其增殖及细胞集落形成。
Objective To investigate the effect of osteoblast precursor cells (MC3T3-E1) on the expression of bone mimicry and proliferation in MDA-MB-231 breast cancer cells by co-culture system. Methods Experimental group MC3T3-E1 and MDA-MB-231 cells were co-cultured in transwell chamber, MC3T3-E1 osteoblasts were inoculated in the upper chamber, and the same number of MDA-MB-231 cells were inoculated in the lower chamber. The same volume and density of MDA-MB-231 cells were used in the upper and lower chambers of the control group. The mRNA and protein expression of osteocalcin (OCN), osteopontin (OPN) and osteoprotegerin (OPG) in experimental group and control group were detected by real-time fluorescence quantitative PCR and Western blot. -8 method to detect differences in the proliferation of each group, crystal violet staining assay cell colony formation differences. Results Compared with the control group, the gene and protein expressions of OCN, OPN and OPG in experimental group were significantly increased (P <0.05), and the proliferation of MDA-MB-231 cells was significantly increased (P <0.01) The number and size of -MB-231 cells colony formation were better than the control group. Conclusion MC3T3-E1 osteoblast can promote the expression of bone mimicry and promote the proliferation and colony formation of breast cancer MDA-MB-231 cells after short-term co-culture.