LINC01048对膀胱癌细胞迁移、侵袭及对细胞外基质受体相互作用通路蛋白和肿瘤肺转移的影响

Effects of LINC01048 on migration, invasion, ECM-receptor interaction pathway proteins, and lung metastasis of bladder cancer cells

摘要:

目的 探讨长链非编码RNA LINC01048对膀胱癌细胞迁移、侵袭及对细胞外基质（ECM）受体相互作用通路蛋白（Collagen I、α-SMA、ITGA6和CD44）和肿瘤肺转移的影响。方法 采用实时荧光定量聚合酶链反应（qRT-PCR）检测正常膀胱上皮细胞系（SV-HUC-1）及膀胱癌细胞系（T24、5637、J82、UM-UC-3）LINC01048基因表达。选取LINC01048基因表达显著上调的T24细胞系进行si-LINC01048转染实验，构建si-LINC01048组和si-NC组，并设置对照组（未转染的T24细胞）。采用MTT法检测细胞活力、划痕实验检测细胞迁移率、Transwell实验检测细胞侵袭数，Western blotting检测Collagen I、α-SMA、ITGA6和CD44蛋白表达，复制裸鼠膀胱癌肺转移模型，采用HE染色观察转染后裸鼠肺组织的肿瘤转移情况。结果 LINC01048在T24、5637、J82、UM-UC-3细胞系中相较于SV-HUC-1细胞系显著上调，其中T24细胞系LINC01048基因表达最高。si-LINC01048转染后，对照组、si-NC组、si-LINC01048组在0、24和48 h的细胞活力比较，采用重复测量设计的方差分析，结果 ①不同时间点的细胞活力比较，差异有统计学意义（P <0.05）；②3组的细胞活力比较，差异有统计学意义（P <0.05）；③3组细胞活力变化趋势比较，差异有统计学意义（P <0.05）。与对照组比较，si-NC组细胞迁移率及细胞侵袭数差异无统计学意义（P >0.05），si-LINC01048组细胞迁移率及细胞侵袭数下降（P <0.05）；与si-NC组比较，si-LINC01048组细胞迁移率及细胞侵袭数下降（P <0.05）。与对照组相比，si-NC组细胞Collagen I、α-SMA、ITGA6和CD44蛋白表达差异无统计学意义（P >0.05），而si-LINC01048组Collagen I、α-SMA、ITGA6和CD44蛋白表达均下降（P <0.05）；与si-NC组比较，si-LINC01048组细胞Collagen I、α-SMA、ITGA6和CD44蛋白表达均下降（P <0.05）。此外，沉默LINC01048能够有效抑制膀胱癌细胞在小鼠肺部的转移（P <0.05）。结论 LINC01048在膀胱癌中呈高表达，沉默LINC01048能显著抑制膀胱癌细胞的活力、迁移、侵袭及转移，提示LINC01048可能作为膀胱癌治疗的潜在靶点。

Abstract:

Objective To investigate the role of long non-coding RNA LINC01048 in the migration, invasion, extracellular matrix (ECM)-receptor interaction pathway proteins (Collagen I, α-SMA, ITGA6, and CD44), and lung metastasis of bladder cancer cells.Methods The expression levels of the LINC01048 gene were detected through qRT-PCR in normal bladder epithelial cells (SV-HUC-1) and bladder cancer cell lines (T24, 5637, J82, UM-UC-3). The T24 cell line, which exhibited the highest expression of LINC01048, was selected for si-LINC01048 transfection experiments to establish an si-LINC01048 group, an si-NC group, and a control group (non-transfected T24 cells). Cell viability was measured using the MTT assay, migration rates by scratch assays, invasion capabilities by Transwell assays, and protein expressions (Collagen I, α-SMA, ITGA6, and CD44) were assessed through Western blotting. Additionally, a nude mouse model of bladder cancer lung metastasis was constructed, and tumor metastasis in lung tissues was evaluated by HE staining.Results LINC01048 was significantly upregulated in bladder cancer cell lines (T24, 5637, J82, UM-UC-3) compared with SV-HUC-1 cells, with the highest expression observed in the T24 cell line. Following si-LINC01048 transfection: (1) The comparison of cell viability at 0, 24, and 48 hours across groups showed significant differences between different time points (P <0.05), across three groups (P <0.05), and in their trends of change (P <0.05). (2) Compared with the control group, the si-NC group showed no significant effect on migration rate or invasion capacity (P >0.05), while the si-LINC01048 group exhibited reduced migration and invasion (P <0.05). Similarly, the si-LINC01048 group demonstrated significantly lower migration and invasion compared to the si-NC group (P <0.05). (3) Protein expression analysis showed no significant differences in Collagen I, α-SMA, ITGA6, and CD44 levels between the si-NC and control groups (P >0.05), whereas these protein levels significantly decreased in the si-LINC01048 group compared to both the control and si-NC groups (P <0.05). Furthermore, silencing LINC01048 effectively suppressed lung metastasis in mice (P <0.05).Conclusion LINC01048 is highly expressed in bladder cancer cells, and its silencing significantly inhibits cell viability, migration, invasion, and metastasis. These findings suggest that LINC01048 may serve as a potential therapeutic target for bladder cancer.