TGF-β1 induced hBMSCs exosomes to secrete miR-424-3p to promote the proliferation and migration of prostate cancer cells

doi:10.3877/cma.j.issn.1674-3253.2024.01.015

Abstract

Abstract:

Objective

To investigate the effect of exosomes derived from TGF-β1-induced osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) on the proliferation, migration, and invasion of prostate cancer (PCa) PC-3 cells.

Methods

The impact of TGF-β1-induced osteogenic differentiation of hBMSCs was assessed using ALP ELISA and Western blot. Exosomes were isolated from cell culture supernatants via ultracentrifugation and characterized using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blot. The expression profile of miRNAs in hBMSCs exosomes following TGF-β1 induction was determined using deep RNA sequencing technology, and the expression level of miRNAs in exosomes was measured using qPCR. Cell proliferation was evaluated using the CCK8 assay, cell migration ability was assessed using the Wound-healing assay, and cell invasion ability was determined using the Trans-well assay.

Results

Compared to the control group, TGF-β1 treatment significantly increased the alkaline phosphatase (ALP) activity on hBMSCs and resulted in a significant up regulation of osteogenic-related factors BMP-2, OCN, and RUNX2 at the protein level, with significant differences (P<0.001). Additionally, it was found that compared to the PC3 cells in hBMSCs group, the proliferation, migration, and invasion abilities of PC3 cells in the TGF-β1-treated hBMSCs group were significantly elevated (P<0.001). Subsequently, exosomes were successfully isolated from the culture supernatants of hBMSCs (hBMSCs_Exo group) and BMSCs following TGF-β1 induction (TGF-β1_hBMSCs_Exo group). Under transmission electron microscopy, typical vesicular structures of exosomes were observed, and exosome-specific proteins such as CD9, CD63, and CD81 were expressed. Notably, the concentration of TGF-β1_hBMSCs_Exo was higher than that of hBMSCs_Exo. Based on miRNA sequencing results, 95 miRNAs were found to be upregulated in TGF-β1_hBMSCs_Exo. The top 5 miRNAs were selected for qPCR validation. The results showed that compared to hBMSCs_Exo, miR-424-3p expression was significantly increased in TGF-β1_hBMSCs_Exo (P<0.001), which was consistent with the miRNA sequencing results. Compared to the miRNA NC group, the proliferation, migration, and invasion abilities of PC3 cells in the miR-424-3p mimic group and TGF-β1_hBMSCs_Exo group were all significantly elevated (P<0.001). Furthermore, it was found that there was no significant difference between the proliferation, migration, and invasion abilities of the miR-424-3p mimic group and TGF-β1_hBMSCs_Exo group.

Conclusion

The miR-424-3p present in hBMSCs exosomes induced by TGF-β1 can significantly enhance the proliferation, migration and invasion abilities of PC3 cells. These findings may provide a new target for individualized treatment of bone metastatic PCa.

Key words: TGF-β1, hBMSCs, miRNA, Prostate cancer, Bone metastasis, Osteogenic-related factors, Exosome

Ruifeng Deng, Lu Cheng, Yulin Zhou, Yuanling Liu, Wencong Jiang, Minyao Jiang, Funeng Jiang, Ming Xi. TGF-β1 induced hBMSCs exosomes to secrete miR-424-3p to promote the proliferation and migration of prostate cancer cells[J]. Chinese Journal of Endourology(Electronic Edition), 2024, 18(01): 82-89.

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URL: https://zhqjmnmwkzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-3253.2024.01.015

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References 32

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