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Updated: Aug 8, 2025
Bone Conditioned Medium: Preparation and Bioassay
Published on: July 8, 2015
Conditioned medium of PC‑3 prostate cancer cells affects microRNA and mRNA profiles in mechanically strained
Zhen Cao1,2,3, Zhixiong Yan1,2, Jiahui Wang1,2
1Department of Biomedical Engineering, College of Biotechnology, Guilin Medical University, Guilin, Guangxi 541199, P.R. China.
Abstract:
Bone is the main site of metastasis from prostate cancer; therefore, it is important to investigate the microRNAs (miRNAs) and mRNA associated with bone metastases from prostate cancer. Since an appropriate mechanical environment is important in the growth of bone, in the present study, the miRNA, mRNA, and long non-coding RNA (lncRNA) profiles of mechanically strained osteoblasts treated with conditioned medium (CM) from PC-3 prostate cancer cells were studied. MC3T3-E1 osteoblastic cells were treated with the CM of PC-3 prostate cancer cells and were simultaneously stimulated with a mechanical tensile strain of 2,500 µε at 0.5 Hz; the osteoblastic differentiation of the MC3T3-E1 cells was then assessed. In addition, the differential expression levels of mRNA, miRNA and lncRNA in MC3T3-E1 cells treated with the CM of PC-3 cells were screened, and some of the miRNAs and mRNAs were verified by reverse transcription-quantitative PCR (RT-qPCR). The signal molecules and signaling pathways associated with osteogenic differentiation were predicted by bioinformatics analysis. The CM of PC-3 prostate cancer cells suppressed osteoblastic differentiation of MC3T3-E1 cells. A total of seven upregulated miRNAs and 12 downregulated miRNAs were selected by sequencing and further verified using RT-qPCR, and related differentially expressed genes (11 upregulated and 12 downregulated genes) were also selected by sequencing and further verified using RT-qPCR; subsequently, according to the enrichment of differentially expressed genes in signaling pathways, nine signaling pathways involved in osteogenic differentiation were screened out. Furthermore, a functional mRNA-miRNA-lncRNA regulatory network was constructed. The differentially expressed miRNAs, mRNAs and lncRNAs may provide a novel signature in bone metastases of prostate cancer. Notably, some of the signaling pathways and related genes may be associated with pathological osteogenic differentiation caused by bone metastasis of prostate cancer.
Insights
Prostate cancer cells suppress bone cell differentiation, altering microRNA (miRNA), mRNA, and long non-coding RNA (lncRNA) profiles. These molecular changes and pathways may indicate bone metastasis in prostate cancer patients.
Area of Science:
- Molecular Biology
- Oncology
- Biochemistry
Background:
- Bone is a primary site for prostate cancer metastasis.
- Understanding the molecular mechanisms of prostate cancer bone metastasis is crucial.
- Mechanical forces influence bone biology and cancer cell interactions.
Purpose of the Study:
- To investigate the molecular profiles (miRNA, mRNA, lncRNA) of osteoblasts under mechanical strain and prostate cancer cell influence.
- To identify signaling pathways involved in prostate cancer-induced bone metastasis.
- To explore potential molecular signatures for prostate cancer bone metastasis.
Main Methods:
- MC3T3-E1 osteoblastic cells were treated with conditioned medium from PC-3 prostate cancer cells.
- Cells were subjected to mechanical tensile strain (2,500 µε at 0.5 Hz).
- Differential expression of mRNA, miRNA, and lncRNA was analyzed using sequencing and verified by RT-qPCR; bioinformatics analysis predicted signaling pathways.
Main Results:
- Prostate cancer conditioned medium suppressed osteoblastic differentiation.
- Sequencing identified seven upregulated and 12 downregulated miRNAs, and 11 upregulated and 12 downregulated mRNAs.
- Nine signaling pathways involved in osteogenic differentiation were identified, and a regulatory network was constructed.
Conclusions:
- Differentially expressed miRNAs, mRNAs, and lncRNAs may serve as novel signatures for prostate cancer bone metastasis.
- Identified signaling pathways and genes are potentially linked to pathological osteogenic differentiation in bone metastasis.
- This study provides insights into the molecular interplay between prostate cancer cells and bone microenvironment.
