乳腺癌细胞条件培养基抑制成骨细胞增殖的作用研究
|
摘要
目的以高转移恶性乳腺癌细胞MDA-MB-231和恶性程度较低的MCF-7乳腺癌细胞为研究对象,探讨两种乳腺癌细胞条件培养基对hFOB1.19细胞增殖作用的影响。方法收集MDA-MB-231、MCF-7和正常对照MCF-10的条件培养基用于hFOB1.19细胞的培养,并采用MTT法检测细胞的增殖效果,流式细胞术检测细胞的生长周期,Western blot检测其周期蛋白CyclinA2的表达。结果以加入不同用量的MDA-MB-231、MCF-7条件培养基培养hFOB1.19细胞和以60%各细胞条件培养基培养hFOB1.19细胞不同时间后,hFOB1.19细胞的增殖均受到抑制,且呈剂量和作用时间依赖性;两者明显影响hFOB1.19细胞的S期,其周期蛋白Cyclin A2的表达在低浓度出现上调,但随浓度的增加而表达下降,而正常对照MCF-10条件培养基对hFOB1.19细胞的增殖影响无显著差异。结论 MDA-MB-231、MCF-7条件培养基对hFOB1.19细胞的增殖具有抑制作用,可能与其影响hFOB1.19细胞周期的S期和Cyclin A2的表达有关。
Objective To study the effects of two kinds of breast cancer cell line conditioned media on the proliferation of hFOB1.19 cells, by the high-metastatic malignant breast cancer cell line MDA-MB-231 and the less malignant breast cancer cell line MCF-7. Methods The conditioned medium of MDA-MB-231, MCF-7 and MCF-10 as a normal control were used to culture hFOB1.19 cells. Cell proliferation was measured by MTT method, cell growth cycle was detected by flow cytometry and the expression of cyclin A2 was detected by western blot(WB). Results After hFOB1.19 cells were cultured in different doses of conditioned medium for different time, the proliferation of hFOB1.19 cell line were inhibited by either MDA-MB-231, MCF-7 conditioned medium or 60% of each cell conditioned medium. Moreover the inhibitory effect was dose-dependent and time-dependent. Both conditioned medium significantly affected the S phase of hFOB1.19 cells and the Cyclin A2 levels of hFOB1.19 cells was up-regulated at low concentration, but down-regulated with the increase of concentration. However the effect of the normal control MCF-10 conditioned medium on the proliferation of hFOB1.19 cell line were no significant differences. Conclusion MDA-MB-231 and MCF-7 conditioned medium may inhibit the proliferation of hFOB1.19 cell line by affecting the expression of S phase and Cyclin A2 levels of hFOB1.19 cells.
Objective To study the effects of two kinds of breast cancer cell line conditioned media on the proliferation of hFOB1.19 cells, by the high-metastatic malignant breast cancer cell line MDA-MB-231 and the less malignant breast cancer cell line MCF-7. Methods The conditioned medium of MDA-MB-231, MCF-7 and MCF-10 as a normal control were used to culture hFOB1.19 cells. Cell proliferation was measured by MTT method, cell growth cycle was detected by flow cytometry and the expression of cyclin A2 was detected by western blot(WB). Results After hFOB1.19 cells were cultured in different doses of conditioned medium for different time, the proliferation of hFOB1.19 cell line were inhibited by either MDA-MB-231, MCF-7 conditioned medium or 60% of each cell conditioned medium. Moreover the inhibitory effect was dose-dependent and time-dependent. Both conditioned medium significantly affected the S phase of hFOB1.19 cells and the Cyclin A2 levels of hFOB1.19 cells was up-regulated at low concentration, but down-regulated with the increase of concentration. However the effect of the normal control MCF-10 conditioned medium on the proliferation of hFOB1.19 cell line were no significant differences. Conclusion MDA-MB-231 and MCF-7 conditioned medium may inhibit the proliferation of hFOB1.19 cell line by affecting the expression of S phase and Cyclin A2 levels of hFOB1.19 cells.
引文
[1] Wang J,Rouse C,Jasper JS,et al.ABL kinases promote breast cancer osteolytic metastasis by modulating tumor-bone interactions through TAZ and STAT5 signaling[J].Sci Signal,2016,9(413):ra12.
[2] Telera S,Caroli F,Raus L,et al.Spine surgery in patients with metastatic breast cancer:A retrospective analysis[J].World Neurosurg,2016,90:133-146.
[3] Wu MY,Li CJ,Yiang GT,et al.Molecular regulation of bone metastasis pathogenesis[J].Cell Physiol Biochem,2018,46(4):1423-1438.
[4] MaoY,Keller ET,Garfield DH,et al.Stroma cells in tumor microenvironment and breast cancer[J].cancer Metastasis Rev,2013,32(0):303-315.
[5] Siclar VA,Mohammad KS,Tompkins DR,et al.Tumor-expressed adrenomedullin accelerates breast cancer bone metastasis[J].Breast Cancer Res,2014,16:458.
[6] Hashimoto K,Ochi H,Sunamura S,et al.Cancer-secreted hsa-miR-940 induces an osteoblastic phenotype in the bone metastatic microenvironment via targeting ARHGAP1 and FAM134A[J].Proc Natl Acad Sci USA,2018,115(9):2204-2209.
[7] McFarlane S,Coulter JA,Tibbits P,et al.CD44 increases the efficiency of distant metastasis of breast cancer[J].Oncotarget,2015,6(13):11465-11476.
[8] Vishal M,Swetha R,Thejaswini G,et al.Role of Runx2 in breast cancer-mediated bone metastasis[J].Int J Biol Macromol,2017,99:608-614.
[9] Krzeszinski JY,Wan Y.New therapeutic targets for cancer bone metastasis[J].Trends Pharmacol Sci,2015,36(6):360-373.
[10] 李海艳,龙小康,张苹,等.乳腺癌骨转移的研究进展[J].肿瘤药学,2017,7 (3):262-268.
[11] Ottewell PD.The role of osteoblasts in bone metastasis[J].J Bone Oncol,2016,5(3):124-127
[12] Mittal S,Brown NJ,Holen I .The breast tumor microenvironment:role in cancer development,progression and response to therapy[J].Expert Rev Mol Diagn,2018,18(3):227-243.
[13] Veronesi F,Borsari V,Sartori M,et al.The use of cell conditioned medium for musculoskeletal tissue regeneration[J].J Cell Physiol,2018,233(6):4423-4442.
[14] Khanmohammadi N,Sameni HR,Mohammadi M,et al.Effect of transplantation of bone marrow stromal cell-conditioned medium on ovarian function,morphology and cell death in cyclophosphamide-treated tats[J].Cell J,2018,20(1):10-18.
[15] Gygli PE,Chang JC,Gokozan HN,et al.Cyclin A2 promotes DNA repair in the brain during both development and aging[J].Aging(Albany NY),2016,8(7):1540-1564.
[16] Yang J,Ren X,Zhang L,et al.Oridonin inhibits oral cancer growth and PI3K/Akt signaling pathway[J].Biomed Pharmacother,2018,100:226-232.
[17] Huang HL,Chen WC,Hsu HP,et al.Argininosuccinate lyase is a potential therapeutic target in breast cancer[J].Oncol Rep,2015,34(6):3131-3139.
[18] Yazdani S,Kasajima A,Onodera Y,et al.Progesterone arrested cell cycle progression through progesterone receptor isoform A in pancreatic neuroendocrine neoplasm[J].J Steroid Biochem Mol Biol,2018,178:243-253.
[19] Loukil A,Cheung CT,Bendris,et al.Cyclin A2:at the crossroads of cell cycle and cell invasion[J].World J Biol Chem,2015,6(4):346-350.
[2] Telera S,Caroli F,Raus L,et al.Spine surgery in patients with metastatic breast cancer:A retrospective analysis[J].World Neurosurg,2016,90:133-146.
[3] Wu MY,Li CJ,Yiang GT,et al.Molecular regulation of bone metastasis pathogenesis[J].Cell Physiol Biochem,2018,46(4):1423-1438.
[4] MaoY,Keller ET,Garfield DH,et al.Stroma cells in tumor microenvironment and breast cancer[J].cancer Metastasis Rev,2013,32(0):303-315.
[5] Siclar VA,Mohammad KS,Tompkins DR,et al.Tumor-expressed adrenomedullin accelerates breast cancer bone metastasis[J].Breast Cancer Res,2014,16:458.
[6] Hashimoto K,Ochi H,Sunamura S,et al.Cancer-secreted hsa-miR-940 induces an osteoblastic phenotype in the bone metastatic microenvironment via targeting ARHGAP1 and FAM134A[J].Proc Natl Acad Sci USA,2018,115(9):2204-2209.
[7] McFarlane S,Coulter JA,Tibbits P,et al.CD44 increases the efficiency of distant metastasis of breast cancer[J].Oncotarget,2015,6(13):11465-11476.
[8] Vishal M,Swetha R,Thejaswini G,et al.Role of Runx2 in breast cancer-mediated bone metastasis[J].Int J Biol Macromol,2017,99:608-614.
[9] Krzeszinski JY,Wan Y.New therapeutic targets for cancer bone metastasis[J].Trends Pharmacol Sci,2015,36(6):360-373.
[10] 李海艳,龙小康,张苹,等.乳腺癌骨转移的研究进展[J].肿瘤药学,2017,7 (3):262-268.
[11] Ottewell PD.The role of osteoblasts in bone metastasis[J].J Bone Oncol,2016,5(3):124-127
[12] Mittal S,Brown NJ,Holen I .The breast tumor microenvironment:role in cancer development,progression and response to therapy[J].Expert Rev Mol Diagn,2018,18(3):227-243.
[13] Veronesi F,Borsari V,Sartori M,et al.The use of cell conditioned medium for musculoskeletal tissue regeneration[J].J Cell Physiol,2018,233(6):4423-4442.
[14] Khanmohammadi N,Sameni HR,Mohammadi M,et al.Effect of transplantation of bone marrow stromal cell-conditioned medium on ovarian function,morphology and cell death in cyclophosphamide-treated tats[J].Cell J,2018,20(1):10-18.
[15] Gygli PE,Chang JC,Gokozan HN,et al.Cyclin A2 promotes DNA repair in the brain during both development and aging[J].Aging(Albany NY),2016,8(7):1540-1564.
[16] Yang J,Ren X,Zhang L,et al.Oridonin inhibits oral cancer growth and PI3K/Akt signaling pathway[J].Biomed Pharmacother,2018,100:226-232.
[17] Huang HL,Chen WC,Hsu HP,et al.Argininosuccinate lyase is a potential therapeutic target in breast cancer[J].Oncol Rep,2015,34(6):3131-3139.
[18] Yazdani S,Kasajima A,Onodera Y,et al.Progesterone arrested cell cycle progression through progesterone receptor isoform A in pancreatic neuroendocrine neoplasm[J].J Steroid Biochem Mol Biol,2018,178:243-253.
[19] Loukil A,Cheung CT,Bendris,et al.Cyclin A2:at the crossroads of cell cycle and cell invasion[J].World J Biol Chem,2015,6(4):346-350.