The mechanism of breast cancer cell culture supernatant promoting migration and proliferation of human adipose mesenchymal stem cells

doi:10.11958/20230255

Abstract

Abstract:

Objective To explore the effect of MDA-MB-231 breast cancer cell culture supernatant on migration, proliferation and apoptosis of human adipose mesenchymal stem cell (hAdMSC) and its molecular mechanism. Methods hAdMSC cultured from MDA-MB-231 cell supernatant and RPMI-1640 medium without fetal bovine serum were mixed at a volume ratio of 1∶4 to form the MDA-MB-231 supernatant group. CXCR1/2 inhibitor group was added with 10 μmol/L Reparixin (CXCR1/2 inhibitor) to the MDA-MB-231 supernatant group. The Akt inhibitor group was added with 10 nmol/L GSK690693 (Akt inhibitor) to the MDA-MB-231 supernatant group. hAdMSC cultured without any stimulation was used as the control group. The migration ability of hAdMSC in each group was detected by cell scratch assay. hAdMSC proliferation was detected by CCK-8 assay. Annexin V-FITC/PI double labeled flow cytometry was used to detect hAdMSC apoptosis rate in each group. The protein expression levels of mTOR/phosphorylated mTOR (p-mTOR) and Akt/phosphorylated Akt (p-Akt) in hAdMSC of each group were detected by Western blot assay. Results Compared with the control group, the 24 h and 48 h scratch closure area, cell proliferation level and relative expression levels of p-Akt, p-mTOR protein of hAdMSC were increased in the MDA-MB-231 supernatant group, and the apoptosis rate was decreased (P<0.05). Compared with the MDA-MB-231 supernatant group, the 48 h cell scratch closure area, cell proliferation level and relative expression levels of p-Akt, p-mTOR proteins of hAdMSC were decreased in the Akt inhibitor group and the CXCR1/2 inhibitor group, and the apoptosis rate was increased (P<0.05). Conclusion MDA-MB-231 breast cancer cell culture supernatant promotes hAdMSC migration and proliferation and inhibits hAdMSC apoptosis by activating Akt-mTOR signaling pathway, in which IL-8-CXCR1/2 axis plays a key role.

Key words: breast neoplasms, cell migration, cell proliferation, tumor microenvironment, adipose derived mesenchymal stem cells

CLC Number:

R737.9

Figures/Tables 7

Tab.1 Comparison of cell scratch closure area of hAdMSC between the four groups

组别	24 h	48 h
对照组	6.57±1.05	20.53±0.69
MDA-MB-231上清液组	39.54±1.98^a	61.19±0.21^a
Akt抑制剂组	35.93±0.48	43.00±0.26^b
CXCR1/2抑制剂组	37.68±0.95	45.23±0.32^b
F	318.300^＊＊	3 240.000^＊＊

Fig.1 Typical images of hAdMSC cell scratches in each group （Scale bar=100 μm）

Fig.2 Histogram of hAdMSC A450 in each group

Fig.3 Histogram of hAdMSC apoptosis rate in each group

Fig.4 Typical images of hAdMSC apoptosis rate detected by Annexin V-FITC/PI double labeled flow cytometry

Fig.5 MDA-MB-231 supernatant promoted expression levels of p-Akt and p-mTOR proteins in hAdMSC

Tab.2 Comparison of the relative expression levels of Akt, p-Akt, mTOR and p-mTOR hAdMSC between the four groups

组别	Akt	p-Akt	mTOR	p-mTOR
对照组	1.11±0.06	0.43±0.02	1.06±0.09	0.59±0.05
MDA-MB-231 上清液组	1.12±0.01	1.03±0.03^a	1.10±0.04	0.94±0.02^a
Akt抑制剂组	1.02±0.05	0.48±0.03^b	1.03±0.02	0.71±0.03^b
CXCR1/2抑制剂组	1.09±0.01	0.77±0.04^b	0.95±0.03	0.73±0.02^b
F	2.622	144.000^＊＊	2.890	86.400^＊＊

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