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目的 :探讨前列腺癌LNCaP和PC-3细胞条件培养液(conditioned medium,CM)对成骨前体细胞MC3T3-E1增殖和分化的影响。方法 :分别用含20%LNCaP CM、20%PC-3 CM和不含CM的α-MEM培养液(作为对照组)处理MC3T3-E1细胞。CCK-8法检测各组细胞的增殖情况,碱性磷酸酶(alkaline phosphatase,ALP)活性检测试剂盒检测各组细胞中ALP的活性,茜素红染色法检测各组细胞的钙化情况,实时荧光定量-PCR法检测各组细胞中Ⅰ型胶原蛋白α1(collagen typeⅠalpha 1,Col1α1)、骨钙蛋白(osteocalcin,OCN)和RUNX2(runt-related transcription factor 2)的mRNA表达水平。结果 :与对照组和LNCaP CM组相比,PC-3CM组MC3T3-E1细胞的增殖能力明显增高(P<0.05);LNCaP CM组MC3T3-E1细胞中ALP的活性明显高于对照组(P<0.05),钙化面积大于对照组(P<0.05);PC-3 CM组MC3T3-E1细胞中ALP的活性较对照组明显降低(P<0.05),钙化面积明显小于对照组(P<0.05);LNCaP CM组MC3T3-E1细胞中Col1α1、OCN和RUNX2的mRNA表达水平均明显高于对照组(P<0.05),而PC-3 CM组MC3T3-E1细胞中Col1α1、OCN和RUNX2的mRNA表达水平均明显低于对照组(P<0.05)。结论 :PC-3 CM可明显促进成骨前体细胞增殖,但抑制其分化成熟;而LNCaP CM可显著促进成骨前体细胞分化成熟,说明该模型能够充分反映不同前列腺癌细胞对成骨前体细胞增殖和分化的差异,可以作为后续研究前列腺癌骨转移机制的体外模型。
Objective: To investigate the effect of conditioned medium (CM) of prostate cancer cell line LNCaP and PC-3 on the proliferation and differentiation of osteoblast precursor cells (MC3T3-E1). Methods: MC3T3-E1 cells were treated with 20% LNCaP CM, 20% PC-3 CM and CM-free α-MEM medium as a control. The proliferation of cells in each group was detected by CCK-8 assay. ALP activity was detected by alkaline phosphatase (ALP) activity assay kit. Alizarin red staining was used to detect the cell calcification and real-time fluorescence The mRNA expression of collagen typeⅠalpha1 (Col1α1), osteocalcin (OCN) and RUNX2 (runt-related transcription factor 2) were detected by quantitative polymerase chain reaction (PCR) Results: The proliferation of MC3T3-E1 cells in PC-3CM group was significantly higher than that in control group and LNCaP CM group (P <0.05). The activity of ALP in MC3T3-E1 cells in LNCaP CM group was significantly higher than that in control group (P < (P <0.05). The calcification area of MC3T3-E1 cells in PC-3 CM group was significantly lower than that in control group (P <0.05), and calcified area was significantly smaller than that in control group (P <0.05) The mRNA expression levels of Col1α1, OCN and RUNX2 in MC3T3-E1 cells in LNCaP CM group were significantly higher than those in control group (P <0.05), while the expression levels of Col1α1, OCN and RUNX2 mRNA in MC3T3-E1 cells in PC-3 CM group were Significantly lower than the control group (P <0.05). CONCLUSION: PC-3 CM can significantly promote the proliferation of osteoblast precursors, but inhibit its differentiation and maturation. However, LNCaP CM can significantly promote the differentiation and maturation of osteoblast precursors, indicating that this model can fully reflect the effect of different prostate cancer cells on osteogenic Differences in somatic cell proliferation and differentiation can be used as in vitro models for the study of bone metastasis of prostate cancer.