PVT1 Promoted the Proliferation and Migration Ability of BMSCs in Glioma C6 Microenvironment

LIU Dong-rong, LIU Yan, WU Pei-lian, ZHENG Lei-lei, HUANG Xiao-ya, MENG Xue-huan, HU Yun

Abstract

To investigate the changes in the proliferation and migration ability of bone marrow mesenchymal stem cells (BMSCs) after indirect co-culturing with glioma C6 cells, and to examine the role of plasmacytoma variant translocation 1 gene (PVT1), a long non-coding RNA (lncRNA), in these changes.  Methods  After separation, cultivation and identification of BMSCs, BMSCs of good growth condition were picked out and indirectly co-cultured with glioma C6 cells in Transwell chambers. These cells are henceforth referred to as the co-culture group. Normal BMSCs cultured separately were the control group. CCK-8 and soft agar colony formation assay were used to examine the proliferation ability of the two groups of cells. Flow cytometry was used to examine the cell cycle. Wound healing assay and Transwell assay were used to explore the migration ability of the cells. Quantitative real-time PCR (qRT-PCR) was used to examine the genetic expression level of PVT1 in the two groups. The above-mentioned tests were repeated after the co-cultured BMSCs were transfected with si-PVT1 (si-PVT1 group) and si-NC (si-NC group). In addition, qRT-PCR was done to evaluate the expression of CyclinD1, a cell cycle protein gene, and matrix metalloproteinases 2 and 9 (MMP2 and MMP9), the migration-related genes in the si-PVT1 and si-NC transfected co-cultured BMSCs.  Results  The BMSCs used in the present study possess the capability of osteogeneic and adipogenic differentiation. Compared with the control group, the co-cultured BMSCs had smaller size, disorderly arrangement and the lack of intercellular contact inhibition. The proliferation and migration ability was significantly enhanced, the proportions of S and G2 phase cells greatly increased and the expression level of PVT1 was significantly up-regulated (P<0.05) in the co-cultured group in comparison with those of the control group. When compared with the si-NC group, the si-PVT1 group showed inhibited proliferation and migration ability of the co-cultured BMSCs; the percentage of G1 phase cells increased, while that of S phase decreased; the expression of PVT1, CyclinD1, MMP2 and MMP9 mRNA also decreased (P<0.05) in the si-PVT1 group.  Conclusion  The enhanced proliferation and migration ability of BMSCs in the glioma C6 microenvironment may be associated with the up-regulated expression of PVT1 .

 

Keywords: Long non-coding RNA (lncRNA), PVT1, Bone marrow mesenchymal stem cells, Indirect co-culture, Proliferation, Migration

 

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References


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