国际肿瘤学杂志

国际肿瘤学杂志 ›› 2026, Vol. 53 ›› Issue (1): 38-46.doi: 10.3760/cma.j.cn371439-20250421-00005

• 论著 • 上一篇    下一篇

ALDH6A1在肾透明细胞癌中的表达及其对肾癌细胞增殖、凋亡和侵袭的影响

张小茜, 程春来()   

  1. 长江航运总医院肿瘤科，武汉 430000
  • 收稿日期:2025-04-21 出版日期:2026-01-08 发布日期:2026-01-13
  • 通讯作者: 程春来，Email： yh1ph7cl@163.com
  • 基金资助:
    武汉市医学科研项目(WX20Q01)

Expression of ALDH6A1 in clear cell renal cell carcinoma and its impacts on proliferation， apoptosis， and invasion of renal cancer cells

Zhang Xiaoxi, Cheng Chunlai()   

  1. Department of Oncology， General Hospital of the Yangtze River Shipping， Wuhan 430000， China
  • Received:2025-04-21 Online:2026-01-08 Published:2026-01-13
  • Supported by:
    Wuhan Medical Scientific Research Program(WX20Q01)

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摘要：

目的 探讨乙醛脱氢酶6家族成员A1（ALDH6A1）在肾透明细胞癌中的表达及其对肾癌细胞增殖、凋亡和侵袭的影响。方法 基于癌症基因组图谱（TCGA）数据库分析ALDH6A1 mRNA在肾透明细胞癌患者和正常人群中表达水平及患者生存时间的差异；收集2021年1月至2024年10月在长江航运总医院治疗的70例肾透明细胞癌患者癌组织及癌旁组织标本，采用反转录实时定量PCR检测ALDH6A1 mRNA表达；免疫组织化学法检测肾透明细胞癌组织中ALDH6A1蛋白表达；分析不同临床病理特征肾透明细胞癌的ALDH6A1蛋白表达情况。体外培养肾癌细胞株786-O，将786-O细胞分为空白对照组（control组）、ALDH6A1过表达组（OE-ALDH6A1组）及阴性对照组（OE-NC组）。采用CCK-8法与EdU实验检测细胞增殖情况；流式细胞术检测细胞凋亡及细胞周期；Transwell小室检测细胞迁移及侵袭能力；蛋白质印迹法检测不同细胞中ALDH6A1、细胞周期蛋白依赖性激酶4（CDK4）、Ki-67、基质金属蛋白酶（MMP）-2、MMP-9、转录因子Twist蛋白表达。结果 TCGA数据库分析发现，相比正常组织[130 368（296 513）]，ALDH6A1 mRNA表达在肾透明细胞癌组织中[15 168（20 826）]显著降低（U=8 946.5，P<0.001）；Kaplan-Meier分析显示，ALDH6A1 mRNA高表达患者（n=421）10年生存率（72.13%）显著高于低表达患者（41.32%，n=112，χ2=8.33，P<0.001）。70例肾透明细胞癌组织及其癌旁组织中ALDH6A1 mRNA相对表达量分别为0.58±0.13、1.03±0.15，差异有统计学意义（t=48.55，P<0.001）；ALDH6A1蛋白阳性表达率分别为32.86%（23/70）、58.57%（41/70），差异有统计学意义（χ2=9.33，P=0.002）；不同病理分级、TNM分期患者癌组织中ALDH6A1表达差异均有统计学意义（χ2=4.51，P=0.034；χ2=6.99，P=0.008）。与人正常肾小管上皮细胞HK-2比较，人肾癌786-O、769-P、ACHN、A498细胞中ALDH6A1 mRNA及蛋白表达均较低（均P<0.05），其中786-O细胞中ALDH6A1 mRNA及蛋白表达最低（均P<0.05）。control组、OE-NC组、OE-ALDH6A1组786-O细胞吸光度（A）值分别为0.65±0.06、0.63±0.05、0.38±0.04，EdU阳性率分别为（48.34±5.21）%、（49.56±5.65）%、（27.34±3.28）%，凋亡率分别为（2.15±0.43）%、（2.32±0.55）%、（33.46±4.36）%，G0/G1期细胞比例分别为（22.46±3.56）%、（23.16±3.72）%、（37.82±5.42）%，S期细胞比例分别为（31.25±3.78）%、（32.89±3.61）%、（26.33±3.87）%，G2/M期细胞比例分别为（46.29±5.15）%、（43.95±5.63）%、（35.85±4.21）%，迁移细胞数分别为（158.46±16.32）、（155.62±15.48）、（84.23±9.65）个，侵袭细胞数分别为（142.35±15.21）、（139.44±13.56）、（72.34±8.61）个，差异均有统计学意义（F=52.91，P<0.001；F=40.22，P<0.001、F=300.05，P<0.001；F=24.23，P<0.001；F=4.96，P=0.022；F=7.11，P=0.007；F=53.15，P<0.001；F=52.16，P<0.001）；与control组和OE-NC组相比，OE-ALDH6A1组细胞A值、EdU阳性率、S期、G2/M期细胞比例、迁移与侵袭细胞数均显著降低（均P<0.05），凋亡率、G0/G1期细胞比例显著升高（均P<0.05）。control组、OE-NC组、OE-ALDH6A1组CDK4、Ki-67、MMP-2、MMP-9、Twist蛋白相对表达量差异均有统计学意义（均P<0.001）；与control组和OE-NC组相比，OE-ALDH6A1组细胞CDK4、Ki-67、MMP-2、MMP-9、Twist蛋白表达均显著降低（均P<0.05）。结论 ALDH6A1在肾透明细胞癌组织与细胞中表达下调，ALDH6A1 mRNA高表达患者预后较好；过表达ALDH6A1可通过调控细胞周期、下调上皮间质转化过程相关蛋白表达，进而抑制肾癌细胞增殖、迁移与侵袭，促进细胞凋亡。

关键词: 癌，肾细胞, 细胞增殖, 细胞凋亡, 肿瘤浸润, 乙醛脱氢酶6家族成员A1

Abstract:

Objective To investigate the expression of aldehyde dehydrogenase 6 family member A1 （ALDH6A1） in clear cell renal cell carcinoma （ccRCC） and its effects on the proliferation， apoptosis， and invasion of renal cancer cells. Methods The Cancer Genome Atlas （TCGA） database was used to analyze the differences in ALDH6A1 mRNA expression levels between ccRCC patients and healthy individuals， as well as the correlation between ALDH6A1 mRNA expression and survival rate of patients. A total of 70 pairs of ccRCC tissues and adjacent tissues were collected from patients who received treatment at General Hospital of the Yangtze River Shipping between January 2021 and October 2024. Reverse transcription-quantitative PCR was performed to detect ALDH6A1 mRNA expression. Immunohistochemistry was used to analyze ALDH6A1 protein expression in ccRCC tissues. The expression of ALDH6A1 protein in different clinicopathological features of ccRCC was analyzed. The renal cancer cell line 786-O was cultured in vitro and divided into blank control group （control group）， ALDH6A1 overexpression group （OE-ALDH6A1 group） and negative control group （OE-NC group）. Cells proliferation was evaluated by CCK-8 assay and EdU assay. Flow cytometry was used to detect cells apoptosis and cells cycle distribution. Transwell chamber assay was used to analyze the migration and invasion abilities of the cells. Western blotting was used to analyze the protein expressions of ALDH6A1， cyclin-dependent kinase 4 （CDK4）， Ki-67， matrix metalloproteinase （MMP）-2， MMP-9， and transcription factor Twist in different cells. Results TCGA database analysis showed that the mRNA expression of ALDH6A1 in ccRCC tissues [15 168 （20 826）] was significantly lower than that in normal tissues [130 368 （296 513）， U=8 946.5， P<0.001]. Kaplan-Meier analysis showed that the 10-year survival rate of patients with high ALDH6A1 mRNA expression （72.13%， n=421） was significantly higher than that of patients with low ALDH6A1 mRNA expression （41.32%， n=112， χ²=8.33， P<0.001）. The relative mRNA expressions of ALDH6A1 in 70 cases of ccRCC tissues and adjacent tissues were 0.58±0.13 and 1.03±0.15， respectively， with a statistically significant difference （t=48.55， P<0.001）. The positive expression rates of ALDH6A1 protein were 32.86% （23/70） and 58.57% （41/70）， respectively， with a statistically significant difference （χ²=9.33， P=0.002）. There were statistically significant differences in the expression of ALDH6A1 in ccRCC tissues among patients with different pathological grades and TNM stages （χ²=4.51， P=0.034； χ²=6.99， P=0.008）. Compared with human normal renal tubular epithelial cells HK-2， the expressions of ALDH6A1 mRNA and protein in human renal cancer cells （786-O， 769-P， ACHN， A498） were all lower （all P<0.05）. Among them， the expression of ALDH6A1 mRNA and protein was the lowest in 786-O cells （all P<0.05）. The absorbance （A） values of 786-O cells in the control group， OE-NC group and OE-ALDH6A1 group were 0.65±0.06， 0.63±0.05 and 0.38±0.04， respectively. The EdU-positive rates were （48.34±5.21）%， （49.56±5.65）% and （27.34±3.28）%， respectively. The apoptosis rates were （2.15±0.43）%， （2.32±0.55）% and （33.46±4.36）%， respectively. The proportions of cells in the G0/G1 phase were （22.46±3.56）%， （23.16±3.72）% and （37.82±5.42）%， respectively. The proportions of cells in the S phase were （31.25±3.78）%， （32.89±3.61）% and （26.33±3.87）%， respectively. The proportions of cells in the G2/M phase were （46.29±5.15）%， （43.95±5.63）% and （35.85±4.21）%， respectively. The numbers of migration cells were 158.46±16.32， 155.62±15.48 and 84.23±9.65， respectively. The numbers of invasion cells were 142.35±15.21， 139.44±13.56 and 72.34±8.61， respectively， all with statistically significant differences （F=52.91， P<0.001； F=40.22， P<0.001； F=300.05， P<0.001； F=24.23， P<0.001； F=4.96， P=0.022； F=7.11， P=0.007； F=53.15， P<0.001； F=52.16， P<0.001）. Compared with the control group and OE-NC group， the A value of cells， the EdU-positive rate， the proportion of cells in the S phase and G2/M phase， and the number of migration and invasion cells in the OE-ALDH6A1 group were significantly decreased （all P<0.05）， while the apoptosis rate and the proportion of cells in G0/G1 phase were significantly increased （all P<0.05）. There were statistically significant differences in the relative expressions of CDK4， Ki-67， MMP-2， MMP-9， and Twist proteins among the control group， the OE-NC group and the OE-ALDH6A1 group （all P<0.001）. Compared with the control group and OE-NC group， the protein expressions of CDK4， Ki-67， MMP-2， MMP-9 and Twist in the OE-ALDH6A1 group were significantly decreased （all P<0.05）. Conclusions ALDH6A1 expression is down-regulated in ccRCC tissues and cells. Patients with high ALDH6A1 mRNA expression have a better prognosis. Overexpression of ALDH6A1 can inhibit the proliferation， migration， and invasion of renal cancer cells， and promote cell apoptosis by regulating the cell cycle and downregulating the expression of proteins related to the epithelial-mesenchymal transition process.

Key words: Carcinoma， renal cell, Cell proliferation, Apoptosis, Neoplasm invasiveness, Aldehyde dehydrogenase 6 family member A1