跨膜蛋白205对结直肠癌患者预后作用的研究

doi:10.3969/j.issn.1673-8640.2026.04.006

摘要/Abstract

摘要：

目的探讨跨膜蛋白205（TMEM205）在结直肠癌（CRC）患者预后评估中的作用。方法基于TCGA数据库分析CRC患者癌组织TMEM205表达与预后的关系。选取2022年9月—2023年10月同济大学医学院附属杨浦医院CRC患者20例，根据1年内是否发生复发分为复发组（10例）、未复发组（10例）。选取检测5种CRC细胞系（HCT116、SW480、Lovo、RKO、HT29）和正常肠上皮细胞系NCM460，构建5-氟尿嘧啶（5-FU）耐药株（HCT116/5-FU、SW480/5-FU）和奥沙利铂（OXA）耐药株（HCT116/OXA、SW480/OXA），并根据耐药株转染siRNA的不同分为敲减组（转染靶向TMEM205的siRNA）和阴性对照组（转染siNC）。采用CCK-8试验、细胞克隆形成试验、免疫印迹法和实时荧光定量聚合酶链反应检测各细胞系、敲减组和阴性对照组的细胞增殖活性、对化疗药物的敏感性以及上皮-间质表型和肿瘤干性标志物的表达情况。提取HCT116亲代细胞、耐药株培养液和CRC患者血清中的外泌体，并检测TMEM205相对表达量。结果基于TCGA数据库的分析结果显示，TMEM205低表达组生存情况优于高表达组[风险比（HR）=1.72，95%可信区间（CI）为1.107~2.659，P=0.016 1]；癌组织TMEM205表达、TNM病理分期和血清癌胚抗原（CEA）水平均与CRC患者预后不良密切相关，TMEM205表达的得分跨度优于血清CEA。5种CRC细胞系TMEM205 mRNA相对表达量和蛋白表达量均显著高于NCM460细胞（P<0.01）。耐药株TMEM205 mRNA相对表达量和蛋白表达量均显著高于亲代细胞。与阴性对照组比较，敲减组对5-FU和OXA的敏感性增加（P<0.001），半数抑制浓度（IC₅₀）值下降（P<0.001），细胞克隆形成能力降低（P<0.001）；上皮表型相关标志物[上皮型钙黏蛋白（E-Cad）、β-连环蛋白（β-catenin）和γ-连环蛋白（γ-catenin）]表达升高（P<0.01），间质表型相关标志物[波形蛋白（Vimentin）、α-平滑肌肌动蛋白（α-SMA）和神经型钙黏蛋白（N-Cad）]表达降低（P<0.01），肿瘤干性标志物[CD44、八聚体结合转录因子4（OCT4）、Nanog同源框蛋白（Nanog）和性别决定区Y框蛋白2（SOX2）]表达降低（P<0.01）。结论在肠癌细胞中，高表达的TMEM205可通过上皮-间质转化和肿瘤干性维持5-FU和OXA的耐药表型。TMEM205可作为CRC预后的高危因素。血清外泌体TMEM205可作为CRC复发潜在的生物标志物。

关键词: 跨膜蛋白205, 外泌体, 5-氟尿嘧啶, 奥沙利铂, 结直肠癌, 复发, 预后

Abstract:

Objective To investigate the prognostic role of transmembrane protein 205（TMEM205） in patients with colorectal cancer（CRC）. Methods The expression level of CRC tissue TMEM205 was analyzed in the Cancer Genome Atlas（TCGA） database in relation to the prognosis. Serum samples from 20 CRC patients at Yangpu Hospital of Tongji University School of Medicine from September 2022 to October 2023 were collected，and the research subjects were classified into recurrence group（10 cases） and non-recurrence group（10 cases） according to 1-year prognosis. Totally，5 CRC cell lines（HCT116，SW480，Lovo，RKO，HT29） and normal intestinal epithelial cell line NCM460 were selected. 5-Fluorouracil（5-FU）-resistant cell lines（HCT116/5-FU，SW480/5-FU） and oxaliplatin（OXA）-resistant cell lines（HCT116/OXA，SW480/OXA） were established. The drug-resistant cell lines were classified into knockdown group（transfected with TMEM205-targeting siRNA） and negative control group（transfected with siNC） according to different transfected siRNA. CCK-8 assay，cell colony formation assay，western blotting and real-time fluorescent quantitative polymerase chain reaction（qRT-PCR） were performed to determine the cell proliferation activity，chemotherapeutic drug sensitivity and the expression levels of epithelial-mesenchymal phenotype and cancer stemness markers in cell lines，knockdown group and negative control group. Exosomes were extracted from the culture media of parental HCT116 cells and drug-resistant cell lines，as well as the serum of CRC patients，and the expression level of TMEM205 was determined. Results Based on the analysis of the TCGA database，the overall survival of patients in low TMEM205 expression group was better than that in high TMEM205 expression group [hazard ratio（HR）=1.72，95% confidence interval（CI）1.107-2.659，P=0.016 1]. In CRC tissues，the expression of TMEM205，TNM pathological stage and serum carcinoembryonic antigen（CEA） level were related to poor prognosis in CRC patients with the score span of TMEM205 expression being superior to that of serum CEA. The TMEM205 relative mRNA and protein expression levels in 5 CRC cell lines were higher than those in NCM460 cell lines（P<0.01）. The TMEM205 relative mRNA and protein expression levels in drug-resistant isolates were higher than those in parental cells. Compared with negative control group，the knockdown group showed increased sensitivity to 5-FU and OXA（P<0.001），decreased half-maximal inhibitory concentration （IC₅₀）（P<0.001），and reduced cell colony formation ability（P<0.001）. The expression levels of epithelial phenotype-related markers [E-cadherin（E-Cad），beta-catenin and gamma-catenin] were enhanced（P<0.01），while the expression levels of mesenchymal phenotype-related markers [Vimentin，alpha-smooth muscle actin（α-SMA） and N-cadherin] were reduced（P<0.01）. The expression levels of tumor stemness markers [CD44，octamer-binding transcription factor 4（OCT4），Nanog homebox protein（Nanog） and sex-determining region Y-box protein 2（SOX2）] were decreased（P<0.01）. Conclusions In colorectal cancer cells，high expression of TMEM205 can maintain the drug-resistant phenotype to 5-FU and OXA through epithelial-mesenchymal transition and tumor stemness. TMEM205 may serve as a high-risk prognostic factor for CRC. Serum exosomal TMEM205 shows potential as a biomarker for predicting CRC recurrence.

Key words: Transmembrane protein 205, Exosome, 5-Fluorouracil, Oxaliplatin, Colorectal cancer, Recurrence, Prognosis

中图分类号:

R446.1

陈学飞, 黄娟, 任圣洁, 闵向阳, 徐子真. 跨膜蛋白205对结直肠癌患者预后作用的研究[J]. 检验医学, 2026, 41(4): 346-354.

CHEN Xuefei, HUANG Juan, REN Shengjie, MIN Xiangyang, XU Zizhen. Research on the prognostic role of transmembrane protein 205 in patients with colorectal cancer[J]. Laboratory Medicine, 2026, 41(4): 346-354.

图/表 16

参考文献 24

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基因名称	正向引物（5'~3'）	反向引物（5'~3'）
TMEM205	TGAGGTACCAGATTCAGCCC	CTCCATCTTGCAGTCCGTTA
GAPDH	GGAGCGAGATCCCTCCAAAAT	GGCTGTTGTCATACTTCTCATGG

基因名称	正向引物（5'~3'）	反向引物（5'~3'）
TMEM205	TGAGGTACCAGATTCAGCCC	CTCCATCTTGCAGTCCGTTA
GAPDH	GGAGCGAGATCCCTCCAAAAT	GGCTGTTGTCATACTTCTCATGG

细胞系	TMEM205 mRNA	TMEM205蛋白
NCM460	1.00±0.29	1.00±0.05
HCT116	3.87±0.60^*	3.81±0.13^*
SW480	3.97±0.42^*	3.89±0.10^*
Lovo	2.43±0.54^*	4.14±0.04^*
RKO	3.52±0.46^*	2.34±0.96^*
HT-29	2.08±0.35^*	1.94±0.06^*

细胞系	TMEM205 mRNA	TMEM205蛋白
NCM460	1.00±0.29	1.00±0.05
HCT116	3.87±0.60^*	3.81±0.13^*
SW480	3.97±0.42^*	3.89±0.10^*
Lovo	2.43±0.54^*	4.14±0.04^*
RKO	3.52±0.46^*	2.34±0.96^*
HT-29	2.08±0.35^*	1.94±0.06^*

细胞系	TMEM205 mRNA	TMEM205蛋白
HCT116	1.00±0.25	1.00±0.07
HCT116/5-FU	2.18±0.28^*	3.67±0.18^*
HCT116/OXA	2.55±0.33^*	2.34±0.16^*
SW480	1.00±0.08	1.32±0.09
SW480/5-FU	2.40±0.41^#	2.48±0.26^#
SW480/OXA	2.78±0.23^#	2.13±0.08^#