Effect of breast cancer cell conditioned medium on biological behavior of bone marrow mesenchymal stem cells

doi:10.11958/20231487

Abstract

Abstract:

Objective To explore effects of MCF-7 breast cancer cell conditioned medium on proliferation, apoptosis and migration of bone marrow mesenchymal stem cells (BMSC) and its molecular mechanism. Methods BMSC cultured under normal environment was used as the control group. BMSC cultured with MCF-7 cell conditioned medium was selected as the MCF-7 conditioned medium group. The MCF-7 conditioned medium group supplemented with 10 nmol/L GSK690693 (Akt inhibitor) was used as the Akt inhibitor group. The MCF-7 conditioned medium group added with 10 μmol/L Reparixin (CXCR1/2 inhibitor) was used as the CXCR1/2 inhibitor group. BMSC proliferation was detected by MTT assay. Annexin V-FITC/PI double labeled flow cytometry was used to detect BMSC apoptosis rate in each group. Transwell cell migration assay was used to detect the migration ability of BMSC in each group. The contents of interleukin (IL)-8 protein in two kinds of cell culture supernatant and MCF-7 cell conditioned medium were detected by enzyme-linked immunosorbent assay. The protein expression levels of protein kinase B (Akt)/phosphorylated Akt (p-Akt) and mammalian target of rapamycin (mTOR)/phosphorylated mTOR (p-mTOR) in BMSC of each group were detected by Western blot assay. Results Compared with the control group, cell proliferation level, migration number and the relative expression levels of p-Akt, p-mTOR protein of BMSC were increased in the MCF-7 conditioned medium group, and the apoptosis rate was decreased (P<0.05). Compared with the MCF-7 conditioned medium group, cell proliferation level, migration number and the relative expression levels of p-Akt, p-mTOR protein of BMSC were decreased in the CXCR1/2 inhibitor group and the Akt inhibitor group, and the apoptosis rate was increased (P<0.05). The contents of IL-8 protein in MCF-7 cell conditioned medium and MCF-7 culture supernatant were higher than that in the BMSC culture supernatant (P<0.05). Conclusion MCF-7 cell conditioned medium promotes BMSC proliferation and migration and inhibits BMSC apoptosis by activating Akt-mTOR signaling pathway, in which IL-8-CXCR1/2 axis plays a key role.

Key words: breast neoplasms, tumor microenvironment, cell proliferation, apoptosis, cell movement, bone marrow mesenchymal stem cell

CLC Number:

R737.9

Figures/Tables 6

References 20

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组别	OD值	凋亡率/%	迁移数目/（个/视野）
对照组	0.73±0.01	8.52±0.45	151.40±9.01
MCF-7条件培养基组	1.01±0.02^a	3.38±0.62^a	290.80±25.31^a
CXCR1/2抑制剂组	0.82±0.02^b	6.15±0.54^b	205.80±23.80^b
Akt抑制剂组	0.77±0.02^b	6.59±0.34^b	169.20±9.43^b
F	119.400^＊＊	36.030^＊＊	44.600^＊＊

组别	OD值	凋亡率/%	迁移数目/（个/视野）
对照组	0.73±0.01	8.52±0.45	151.40±9.01
MCF-7条件培养基组	1.01±0.02^a	3.38±0.62^a	290.80±25.31^a
CXCR1/2抑制剂组	0.82±0.02^b	6.15±0.54^b	205.80±23.80^b
Akt抑制剂组	0.77±0.02^b	6.59±0.34^b	169.20±9.43^b
F	119.400^＊＊	36.030^＊＊	44.600^＊＊

组别	Akt	p-Akt	mTOR	p-mTOR
对照组	0.85±0.02	0.54±0.04	0.84±0.03	0.50±0.09
MCF-7条件培养基组	0.91±0.02	0.89±0.01^a	0.89±0.02	0.86±0.04^a
CXCR1/2抑制剂组	0.86±0.01	0.76±0.04^b	0.86±0.03	0.60±0.02^b
Akt抑制剂组	0.89±0.06	0.49±0.02^b	0.88±0.01	0.48±0.02^b
F	1.569	88.590^＊＊	1.957	22.820^＊＊

组别	Akt	p-Akt	mTOR	p-mTOR
对照组	0.85±0.02	0.54±0.04	0.84±0.03	0.50±0.09
MCF-7条件培养基组	0.91±0.02	0.89±0.01^a	0.89±0.02	0.86±0.04^a
CXCR1/2抑制剂组	0.86±0.01	0.76±0.04^b	0.86±0.03	0.60±0.02^b
Akt抑制剂组	0.89±0.06	0.49±0.02^b	0.88±0.01	0.48±0.02^b
F	1.569	88.590^＊＊	1.957	22.820^＊＊