Weng Xu, Li Jin-Song, Fan Song
Dept. of Stomatology, Shantou Central Hospital, Shantou 515031, China.
Dept. of Stomatology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2020 Oct 1;38(5):550-557. doi: 10.7518/hxkq.2020.05.014.
To investigate the mechanism underlying the regulation of the invasion and metastasis of oral squamous cell carcinoma (OSCC) by long-chain noncoding RNA (lncRNA) PCGEM1 through the transforming growth factor (TGF) β2/Smad2 signaling pathways.
A total of 60 OSCC cases were collected. Cancer tissues and normal tissues more than 2 cm away from cancer tissues were also collected. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-148a and lncRNA PCGEM1 in OSCC, adjacent normal tissues, oral mucosa epithelial cells, KB, BcaCD885, SCC-4, CAL27, and SCC-15. The relationship between the expression of lncRNA PCGEM1 and miR-148a and the clinicopathological information of patients was analyzed. The lncRNA PCGEM1-silenced cell line KB-siPCGEM1 and negative control (KB-NC) group were constructed, and KB was used as the blank control group. The effects of lncRNA PCGEM1 on the proliferation, invasion, and migration of KB cells were determined via MTT, Transwell, and scratch assays. The bioinformatics website starBase was used to predict the complementary binding microRNA (miRNA) of lncRNA PCGEM1. Furthermore, the genes that the miRNA could target and bind were predicted in accordance with the website www.microRNA.org. Western blotting analysis was used to detect the expression of TGF β2/Smad2 signaling pathway proteins.
qRT-PCR results showed that the expression level of lncRNA PCGEM1 and miR-148a in OSCC tissues was higher than that in normal tissues (P<0.05). The expression of lncRNA PCGEM1 and miR-148a in the cancer tissues of patients with different TNM grades, lymph node metastasis, and tissue differentiation was statistically significant (P<0.05). Compared with those in the blank control group and the KB-NC group, OD492 nm value was significantly decreased and cell mobility was significantly reduced in the KB-siPCGEM1 group (P<0.05). Bioinformatics predictions showed that lncRNA PCGEM1 could bind to miR-148a in a complementary manner and that miR-148a had a targeted binding site with TGF β2. qRT-PCR and Western blotting analysis results showed that the expression levels of miR-148a, TGF β2, and p-Smad2 in the KB-siPCGEM1 group were significantly lower than those in the blank control and KB-NC groups (P<0.05), and no statistically significant difference between the blank control group and the KB-NC group was observed (P>0.05).
LncRNA PCGEM1 is highly expressed in OSCC. The high expression of lncRNA PCGEM1 may enhance the TGF β2/Smad2 signaling pathway by upregulating miR-148a, thus promoting the development of OSCC.
探讨长链非编码RNA（lncRNA）PCGEM1通过转化生长因子（TGF）β2/Smad2信号通路调控口腔鳞状细胞癌（OSCC）侵袭和转移的机制。
收集60例OSCC病例，同时收集癌组织及距癌组织2 cm以外的正常组织。采用实时定量聚合酶链反应（qRT-PCR）检测miR-148a和lncRNA PCGEM1在OSCC组织、癌旁正常组织、口腔黏膜上皮细胞、KB、BcaCD885、SCC-4、CAL27和SCC-15中的表达。分析lncRNA PCGEM1和miR-148a的表达与患者临床病理信息的关系。构建lncRNA PCGEM1沉默细胞系KB-siPCGEM1和阴性对照组（KB-NC），以KB作为空白对照组。通过MTT法、Transwell法和划痕实验检测lncRNA PCGEM1对KB细胞增殖、侵袭和迁移的影响。利用生物信息学网站starBase预测lncRNA PCGEM1的互补结合微小RNA（miRNA），并根据网站www.microRNA.org预测该miRNA可能靶向结合的基因。采用蛋白质印迹法检测TGF β2/Smad2信号通路蛋白的表达。
qRT-PCR结果显示，lncRNA PCGEM1和miR-148a在OSCC组织中的表达水平高于正常组织（P<0.05）。lncRNA PCGEM1和miR-148a在不同TNM分级、有淋巴结转移及组织分化程度的患者癌组织中的表达差异有统计学意义（P<0.05）。与空白对照组和KB-NC组相比，KB-siPCGEM1组的OD492 nm值显著降低，细胞迁移能力显著降低（P<0.05）。生物信息学预测显示，lncRNA PCGEM1可与miR-148a互补结合，且miR-148a与TGF β2有靶向结合位点。qRT-PCR和蛋白质印迹法分析结果显示，KB-siPCGEM1组中miR-148a、TGF β2和p-Smad2的表达水平显著低于空白对照组和KB-NC组（P<0.05），空白对照组和KB-NC组之间差异无统计学意义（P>0.05）。
lncRNA PCGEM1在OSCC中高表达。lncRNA PCGEM1的高表达可能通过上调miR-148a增强TGF β2/Smad2信号通路，从而促进OSCC的发展。
