Yang Jiangeng, Xia An, Zhang Huajie, Liu Qi, You Hongke, Ding Daoyuan, Yin Yonghua, Wen Bo
Department of Urology, Shenzhen Hospital of Integrated Traditional Chinese and Western Medicine, Shenzhen, China.
Front Mol Biosci. 2021 Mar 29;8:654718. doi: 10.3389/fmolb.2021.654718. eCollection 2021.
LncRNAs are defined as non-coding RNAs that are longer than 200 nucleotides in length. The previous studys has shown that lncRNAs played important roles in the regulation of gene expression and were essential in mammalian development and disease processes. Inspired by the observation that lncRNAs are aberrantly expressed in tumors, we extracted RNA from Bladder urothelial carcinoma and matched histologically normal urothelium from each patient and bladder carcinoma cell lines. Then, we reversed transcribed them into cDNA.Last, we investigated the expression patterns of ERIC by the fluorescence quantitative PCR in bladder cancer tissues and cell lines. CRISPR-dCas9-VPR targeting ERIC plasmid was transfected into T24 and 5637 cells, and cells were classified into two groups: negative control (NC) and ERIC overexpression group. MTT assay, transwell assay, and flow cytometry were performed to examine changes in cell proliferation, invasiveness, and apoptosis. We found that the expression of ERIC was down-regulated in bladder urothelial carcinoma compared to matched histologically normal urotheliam. The differences of the expression of this gene were large in the bladder cancer lines. Compared with the negative control group, the ERIC overexpression group showed significantly decreased cell proliferation rate ( = 7.583, = 0.002; = 3.283, 0.03) and invasiveness ( = 11.538, < 0.001; = 8.205, 0.01); and increased apoptotic rate ( = -34.083, < 0.001; = -14.316, < 0.001). Our study lays a foundation for further study of its pathogenic mechanism in bladder cancer.
长链非编码RNA（lncRNAs）被定义为长度超过200个核苷酸的非编码RNA。先前的研究表明，lncRNAs在基因表达调控中发挥重要作用，并且在哺乳动物发育和疾病过程中至关重要。受lncRNAs在肿瘤中异常表达这一观察结果的启发，我们从膀胱尿路上皮癌中提取RNA，并从每位患者和膀胱癌细胞系中获取组织学上正常的尿路上皮作为对照。然后，我们将它们逆转录成cDNA。最后，我们通过荧光定量PCR研究膀胱癌组织和细胞系中ERIC的表达模式。将靶向ERIC质粒的CRISPR-dCas9-VPR转染到T24和5637细胞中，并将细胞分为两组：阴性对照组（NC）和ERIC过表达组。进行MTT试验、Transwell试验和流式细胞术以检测细胞增殖、侵袭和凋亡的变化。我们发现，与匹配的组织学上正常的尿路上皮相比，膀胱尿路上皮癌中ERIC的表达下调。该基因在膀胱癌细胞系中的表达差异很大。与阴性对照组相比，ERIC过表达组的细胞增殖率显著降低（T24：t = 7.583，P = 0.002；5637：t = 3.283，P = 0.03）和侵袭性显著降低（T24：t = 11.538，P < 0.001；5637：t = 8.205，P = 0.01）；凋亡率增加（T24：t = -34.083，P < 0.001；5637：t = -14.316，P < 0.001）。我们的研究为进一步研究其在膀胱癌中的致病机制奠定了基础。
