[前列腺癌细胞系PC-3条件培养基促进人骨髓间充质干细胞的增殖和成骨分化]

[Prostate cancer cell line PC-3 conditioned medium promotes proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells].

作者信息

Tang Yuan-jie, Sun Ying-hao, Xu Chuan-liang, Wang Lin-hui, Gao Xu, Liu Bing, Ji Jia-tao

机构信息

Department of Urology, Changhai Hospital, The Second Military Medical University, Shanghai 200433, China.

出版信息

Zhonghua Nan Ke Xue. 2011 Mar;17(3):229-36.

PMID:21485544

Abstract

OBJECTIVE

To investigate the effects of prostate cancer cell line PC-3 conditioned medium (PC- 3-CM) on the proliferation and osteogenic differentiation of human bone marrow human basalis mesenchymal stem cells (hBMSCs).

METHODS

hBMSCs were isolated and culture-expanded by density gradient centrifugation from normal volunteers. PC-3 cells were cultured till the time of logarithmic growth and then transferred to a fresh medium, which, after 24 hours of incubation, was collected as PC-3-CM. Passage 3 hBMSCs were cultured in the fresh medium alone (the control group) or that with 50% PC-3-CM (the experimental group), and the effect of PC-3-CM on the proliferation activity of the hBMSCs was detected by WST-8 assay. Based on the types of medium used, the hBMSCs were divided into Groups I (control), II (50% PC-3-CM), III (osteoblast inducer) and IV (osteoblast inducer containing 50% PC-3 CM). The effects of PC-3-CM on the osteoblastic differentiation of the hBMSCs were determined by ALP staining, ALP activity detection, Von Kossa staining, and calcium quantitation.

RESULTS

At 1, 3, 5 and 7 days of incubation, the absorbance values of the cells in the experimental group were 0.4370 +/- 0.0285, 0.7980 +/- 0.0213, 1.9090 +/- 0.0612 and 2.3023 +/- 0.0610, and those in the control group were 0.4060 +/- 0.0223, 0.6643 +/- 0.0075, 1.3727 +/- 0.0176 and 1.7947 +/- 0.0115, respectively, with significant differences between the two groups (P < 0.01) except on day 1 (P > 0.05). The positive rate and intensity of ALP staining were gradually increased in the four groups, with the ALP activities of 0.29 +/- 0.03, 1.30 +/- 0.03, 2.13 +/- 0.08, and 3.80 +/- 0.03, respectively (P < 0.01), and so was the intensity of Von Kossa staining, with the calcium depositions of 0.04 +/- 0.01, 0.44 +/- 0.05, 0.98 +/- 0.03, and 1.27 +/- 0.04, respectively (P < 0.01).

CONCLUSION

PC-3- CM can promote the proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells.

摘要

目的

探讨前列腺癌细胞系PC-3条件培养基（PC-3-CM）对人骨髓基底间充质干细胞（hBMSCs）增殖和成骨分化的影响。

方法

通过密度梯度离心法从正常志愿者中分离并培养扩增hBMSCs。培养PC-3细胞至对数生长期，然后转移至新鲜培养基中，孵育24小时后收集作为PC-3-CM。第3代hBMSCs分别培养于单纯新鲜培养基（对照组）或含50%PC-3-CM的新鲜培养基（实验组）中，采用WST-8法检测PC-3-CM对hBMSCs增殖活性的影响。根据所用培养基类型，将hBMSCs分为I组（对照组）、II组（50%PC-3-CM组）、III组（成骨诱导剂组）和IV组（含50%PC-3-CM的成骨诱导剂组）。通过碱性磷酸酶（ALP）染色、ALP活性检测、Von Kossa染色和钙定量测定来确定PC-3-CM对hBMSCs成骨分化的影响。

结果

孵育1、3、5和7天时，实验组细胞的吸光度值分别为0.4370±0.0285、0.7980±0.0213、1.9090±0.0612和2.3023±0.0610，对照组细胞的吸光度值分别为0.4060±0.0223、0.6643±0.0075、1.3727±0.0176和1.7947±0.0115，除第1天外两组间差异有统计学意义（P<0.01）（第1天P>0.05）。四组中ALP染色的阳性率和强度逐渐增加，ALP活性分别为0.29±0.03、1.30±0.03、2.13±0.08和3.80±0.03（P<0.01），Von Kossa染色强度也如此，钙沉积量分别为0.04±0.01、0.44±0.05、0.98±0.03和1.27±0.04（P<0.01）。