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PC-3前列腺癌细胞的条件培养基影响机械拉伸成骨细胞中的微小RNA和信使核糖核酸谱。

Conditioned medium of PC‑3 prostate cancer cells affects microRNA and mRNA profiles in mechanically strained osteoblasts.

作者信息

Cao Zhen, Yan Zhixiong, Wang Jiahui, Yang Huan, Han Biao, Gao Jintao, Guo Yong

机构信息

Department of Biomedical Engineering, College of Biotechnology, Guilin Medical University, Guilin, Guangxi 541199, P.R. China.

Key Laboratory of Biochemistry and Molecular Biology, Guilin Medical University, Education Department of Guangxi Zhuang Autonomous Region, Guilin, Guangxi 541199, P.R. China.

出版信息

Exp Ther Med. 2023 Feb 13;25(3):138. doi: 10.3892/etm.2023.11837. eCollection 2023 Mar.

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.

摘要

骨是前列腺癌转移的主要部位;因此,研究与前列腺癌骨转移相关的微小RNA(miRNA)和信使核糖核酸(mRNA)具有重要意义。由于适宜的力学环境对骨的生长至关重要,在本研究中,对用PC-3前列腺癌细胞的条件培养基(CM)处理的机械拉伸应变成骨细胞的miRNA、mRNA和长链非编码RNA(lncRNA)谱进行了研究。用PC-3前列腺癌细胞的CM处理MC3T3-E1成骨细胞,并同时以0.5 Hz的频率施加2500 με的机械拉伸应变;然后评估MC3T3-E1细胞的成骨分化情况。此外,筛选了用PC-3细胞的CM处理的MC3T3-E1细胞中mRNA、miRNA和lncRNA的差异表达水平,并用逆转录定量聚合酶链反应(RT-qPCR)对部分miRNA和mRNA进行了验证。通过生物信息学分析预测了与成骨分化相关的信号分子和信号通路。PC-3前列腺癌细胞的CM抑制了MC3T3-E1细胞的成骨分化。通过测序选择了总共7个上调的miRNA和l2个下调的miRNA,并使用RT-qPCR进一步验证,同时通过测序选择了相关的差异表达基因(11个上调基因和12个下调基因),并使用RT-qPCR进一步验证;随后,根据差异表达基因在信号通路中的富集情况,筛选出9条参与成骨分化的信号通路。此外,构建了一个功能性mRNA-miRNA-lncRNA调控网络。差异表达的miRNA、mRNA和lncRNA可能为前列腺癌骨转移提供一种新的标志物。值得注意的是,一些信号通路和相关基因可能与前列腺癌骨转移引起的病理性成骨分化有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/59ee/9947580/b7d3ad0739cc/etm-25-03-11837-g00.jpg

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