MTP18 promotes oral squamous-cell carcinoma metastasis through induction of epithelial-mesenchymal transition
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摘要:
目的 探讨线粒体蛋白18(mitochondrial protein 18，MTP18)在口腔鳞癌转移中的调控作用。 方法 (1) 采用生物信息分析44例正常口腔黏膜与520例口腔鳞癌组织中MTP18的表达。(2)siRNA干涉MTP18表达后，用qRT-PCR与Western blotting实验检测干涉效果，用划痕与Transwell实验检测对细胞迁移与侵袭能力的影响。(3)siRNA下调MTP18表达后，用qRT-PCR实验分析对细胞上皮间质转化的影响。 结果 (1) 口腔鳞癌中MTP18表达显著上调(P＜0.001)，尤其在高级别与淋巴转移的肿瘤组织中。(2)siRNA显著下调了MTP18表达(P＜0.001)。干涉MTP18可显著抑制口腔鳞癌SCC9细胞的迁移(siCtrl vs.siMTP18#1 vs. siMTP18#2为1.00±0.07 vs. 0.43±0.04 vs. 0.44±0.03，F=102.241, P＜0.001)与侵袭(siCtrl vs. siMTP18#1 vs. siMTP18#2为1.00±0.07 vs. 0.43±0.04 vs. 0.44±0.03，F=97.017, P＜0.001)。(3)干涉MTP18上调了上皮细胞标志分子的表达(P＜0.001)，而下调了间质细胞标志分子的表达(P＜0.001)。 结论 MTP18通过诱导上皮间质转化促进口腔鳞癌细胞的迁移与侵袭。 Abstract:Objective To explore the biological functions of mitochondrial protein 18 (MTP18) in oral squamous-cell carcinoma metastasis. Methods (1) Bioinformatic analysis was conducted to evaluate the expression of MTP18 in 44 normal oral mucosa and 520 oral squamous-cell carcinoma tissues. (2) The silencing effect of MTP18 by siRNA was measured by qRT-PCR and Western blot analysis. In addition, the effects of MTP18 knockdown on the migration and invasion abilities of oral squamous-cell carcinoma were determined by wound-healing and Transwell invasion assays. (3) The effects of MTP18 knockdown on the expressions of markers in epithelial-mesenchymal transition (EMT) were determined by qRT-PCR analysis. Results (1) Bioinformatic analysis indicated that MTP18 expression was significantly increased in tumour tissues of oral squamous-cell carcinoma, especially in tumour tissues at high stages and with lymph node metastasis, compared with normal tissues (P < 0.001). (2) siRNA transfection successfully reduced the expression of MTP18 in oral squamous-cell carcinoma cells (P < 0.001). MTP18 knockdown significantly suppressed cell migration (siCtrl vs. siMTP18#1 vs. siMTP18#2: 1.00±0.07 vs. 0.43±0.04 vs. 0.44±0.03, F=102.241, P < 0.001) and invasion (siCtrl vs. siMTP18#1 vs. siMTP18#2: 1.00±0.07 vs. 0.43±0.04 vs. 0.44±0.03, F=97.017, P < 0.001). (3) MTP18 knockdown significantly increased the expressions of epithelial cell markers (P < 0.001), but suppressed the expressions of mesenchymal cell markers (P < 0.001). Conclusion MTP18 overexpression promotes oral squamous-cell carcinoma metastasis mainly through the induction of EMT. -
Key words:
- Mitochondrial protein 18 /
- Mitochondrial fission /
- Migration /
- Invasion /
- Oral squamous-cell carcinoma
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图 1 生物信息分析MTP18在口腔鳞癌中的表达变化
注：A为MTP18在癌组织与正常组织中表达；与正常组比较，aP < 0.001。B为MTP18在不同分级肿瘤中的表达；与正常组比较，aP < 0.001；与肿瘤分级为1级组比较，bP < 0.001；与肿瘤分级为2级组比较，cP < 0.001。C为MTP18在不同分期肿瘤中的表达；与正常组比较，aP < 0.001；与肿瘤分期为1期组比较，bP < 0.001。D为MTP18在转移性与非转移性肿瘤中的表达；与正常组比较，aP < 0.001。
图 2 siRNA对口腔鳞癌SCC9细胞中MTP18表达的干涉效率分析
注：A为qRT-PCR；B为Western blotting。与siCtrl组比较，aP < 0.001。
图 3 划痕实验分析下调MTP18对口腔鳞癌SCC9细胞迁移能力的影响
注：A为典型细胞划痕与迁移图片；B为各组细胞相对迁移距离的统计分析；与siCtrl组比较，aP < 0.001。
图 4 Transwell侵袭实验分析下调MTP18对口腔鳞癌SCC9细胞侵袭能力的影响
注：A为典型细胞侵袭图片；B为各组发生侵袭细胞数的统计分析；与siCtrl组比较，aP < 0.001。
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