| 摘要: |
| 目的 检测翻译控制肿瘤蛋白1（TPT1）在肾细胞癌（RCC）中的表达，并探讨其在RCC中的作用。方法 从基因表达综合数据库下载微阵列数据集GSE15641（包括23例正常肾脏组织样本数据和32例RCC组织样本数据），并使用R 4.3.0软件筛选RCC组织和正常肾脏组织之间的差异表达基因。采用qPCR和蛋白质印迹法检测90例在南通大学附属医院确诊并接受治疗的RCC患者的RCC组织和癌旁正常组织，以及人RCC细胞（769-P、786-O、ACHN和Caki-1）和人正常胚肾293细胞（HEK293）中TPT1的表达。通过χ2检验分析TPT1表达与RCC患者临床病理特征之间的关系。从癌症基因组图谱数据库获得522例RCC患者的临床信息，采用ROC曲线和Kaplan-Meier生存曲线分析TPT1表达与患者预后的相关性。体外转染TPT1 siRNA和siRNA阴性对照（NC）至786-O和Caki-1细胞后，采用qPCR和蛋白质印迹法检测细胞中TPT1的表达，分别应用CCK-8实验、划痕实验和Transwell实验检测细胞的增殖、迁移和侵袭能力，并采用蛋白质印迹法检测凋亡相关蛋白的表达。结果 与正常肾脏组织和HEK293细胞相比，TPT1在RCC组织和RCC细胞中的表达均上调（均P＜0.05）。TPT1表达水平低的RCC患者肿瘤大小及转移发生率均低于TPT1表达水平高的患者（均P＜0.05）。ROC曲线分析结果提示，TPT1对RCC具有较高的诊断价值（AUC=0.856 9，95% CI 0.804 5～0.909 3，P＜0.001）。Kaplan-Meier生存曲线分析结果显示，低TPT1表达组RCC患者的总生存期长于高TPT1表达组（P=0.018 4）。在细胞实验中，与siRNA NC组相比，转染TPT1 siRNA后786-O和Caki-1细胞的增殖活性、划痕愈合率和侵袭穿膜细胞数均下降（均P＜0.01），凋亡相关蛋白Bcl-2和基质金属蛋白酶9表达降低、Bcl-2相关X蛋白表达增加（均P＜0.05）。结论 TPT1参与了RCC的进展，可能是RCC治疗的潜在靶点。 |
| 关键词: 肾细胞癌 翻译控制肿瘤蛋白1 细胞增殖 细胞迁移 细胞凋亡 |
| DOI：10.16781/j.CN31-2187/R.20220585 |
| 投稿时间：2022-07-13修订日期：2022-12-12 |
| 基金项目: |
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| Mechanism of translationally-controlled tumor protein 1 in regulating proliferation and migration of renal cell carcinoma cells |
| WANG Qingqing1,HUANG Xinzhong2*,HU Yinyu3,CAO Jie3 |
(1. Medical School of Nantong University, Nantong 226001, Jiangsu, China;
2. Department of Nephrology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu, China;
3. Department of Nephrology, The Six People's Hospital of Nantong, Nantong 226000, Jiangsu, China
*Corresponding author) |
| Abstract: |
| Objective To detect the expression of translationally-controlled tumor protein 1 (TPT1) in renal cell carcinoma (RCC) and to explore its role in RCC. Methods Microarray dataset GSE15641 (including 23 normal kidney tissue samples and 32 RCC tissue samples) was downloaded from the Gene Expression Omnibus, and differentially expressed genes between RCC tissue and normal kidney tissue were screened using R 4.3.0 software.The TPT1 expression in RCC tissue and adjacent non-tumor tissue of 90 patients diagnosed and treated at Affiliated Hospital of Nantong University, as well as in human RCC cells (769-P, 786-O, ACHN, and Caki-1) and human embryonic kidney 293 cell (HEK293) was detected by quantitative polymerase chain reaction (qPCR) and Western blotting.The relationship between TPT1 expression and clinical pathological characteristics of RCC patients was analyzed by χ2 test.The clinical data of 522 RCC patients were derived from The Cancer Genome Atlas, and the correlation between TPT1 expression and prognosis of RCC patients was analyzed by receiver operating characteristic (ROC) curve and Kaplan-Meier survival curve.After in vitro transfection of TPT1 small interfering RNA (siRNA) and its negative control (NC) into 786-O and Caki-1 cells, the TPT1 expression was detected by qPCR and Western blotting; the proliferation, migration, and invasion were detected by cell counting kit 8 assay, scratch assay, and Transwell assay, respectively; and the expression of apoptosis-related proteins was detected by Western blotting. Results TPT1 expression was significantly upregulated in the RCC tissue and cells compared with the normal kidney tissue and HEK293 cells (all P＜0.05).The RCC patients with low TPT1 expression levels had significantly smaller tumor size and lower metastasis rate than those with high TPT1 expression levels (both P＜0.05).The ROC curve analysis results indicated that TPT1 had high diagnostic value for RCC (area under curve was 0.856 9, 95% confidence interval was 0.804 5-0.909 3, P＜0.001).The Kaplan-Meier survival analysis results showed that the overall survival of RCC patients in the low TPT1 expression group was significantly longer than that in the high TPT1 expression group (P=0.018 4).In the cell experiment, compared with the siRNA NC group, the proliferation activity, scratch healing rate, and invading transmembrane cell number of 786-O and Caki-1 cells were significantly decreased after transfection with TPT1 siRNA (all P＜0.01); the expression levels of B-lymphoma gene 2 (Bcl-2) and matrix metalloproteinase-9 were significantly decreased, while the expression of Bcl-2 associated X protein was significantly increased (all P＜0.05). Conclusion TPT1 is involved in the progression of RCC and may be a potential therapeutic target for RCC. |
| Key words: renal cell carcinoma translationally-controlled tumor protein 1 cell proliferation cell migration apoptosis |
