中国普外基础与临床杂志

中国普外基础与临床杂志

HIBADH 蛋白表达在胃腺癌中的临床意义及其在胃癌细胞中的功能研究

查看全文

目的 研究 3-羟异丁酸脱氢酶(3-hydroxyisobutyrate dehydrogenase,HIBADH)蛋白在胃腺癌中表达的临床意义及其在胃癌细胞中的生物学功能。 方法 回顾收集 2006 年 1 月 1 日至 2007 年 12 月 31 日期间上海交通大学医学院附属第九人民医院普外科收治的 76 例胃腺癌患者。采用免疫组织化学染色检测胃癌原发灶、癌旁组织及癌转移淋巴结中 HIBADH 蛋白的表达,分析胃癌组织中 HIBADH 蛋白的表达与患者临床病理学特征的关系。在细胞层面,采用分子克隆技术构建 HIBADH 稳定敲除的 MKN45 胃癌细胞系。通过细胞增殖实验、迁移实验、侵袭实验及伤口愈合实验(细胞分为转染组和阴性对照组,分别转染干扰序列和阴性对照序列),探索 HIBADH 蛋白在胃癌细胞中的生物学功能。 结果 胃腺癌原发灶组织中的 HIBADH 蛋白表达阳性率较癌旁组织高(χ2=54.738,P<0.001),且胃癌组织中 HIBADH 蛋白的表达与肿瘤直径、淋巴管侵犯、pT 分期、淋巴结转移及 pTNM 分期均相关(P<0.05)。在癌转移的淋巴结中,HIBADH 蛋白的表达阳性率为 100%(48/48)。HIBADH 蛋白表达阳性组和 HIBADH 蛋白表达阴性组患者的术后 10 年生存率分别为 16.4% 和 69.4%,后者的生存时间较长(χ2=19.612,P<0.001)。HIBADH 蛋白表达干扰后,MKN45 胃癌细胞的迁移能力、侵袭能力及伤口愈合能力较阴性对照组均明显下降(P<0.005),但细胞增殖能力无明显变化(P>0.05)。 结论 HIBADH 蛋白在胃癌原发灶中的表达提示肿瘤分期晚、患者预后差。抑制 HIBADH 蛋白表达可降低胃癌细胞的运动能力。

Objective To study the clinical significance of the 3-hydroxyisobutyrate dehydrogenase (HIBADH) protein expression in gastric adenocarcinoma and its biological function in GC cells. Methods Seventy-six patients with GC who were hospitalized in Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiaotong University, School of Medicine between January 1 2006 and December 31 2007 were recruited in our research. Immunohistochemical (IHC) staining was used to detect the HIBADH protein in primary gastric adenocarcinoma tissues, adjacent tissues, and metastatic lymph nodes of gastric cancer. Then, the relationships among the expression of HIBADH protein, the clinical features, and the prognosis were analyzed. The MKN45 gastric cancer cell line of HIBADH overexpression was picked up and constructed as stable HIBADH knockdown cell lines. The biological function of HIBADH protein in gastric cancer cells was confirmed through in vitro experiments such as cell proliferation assay, migration and invasion assay, and scratch-wound assay. Results The positive expression rate of HIBADH protein in the 76 gastric adenocarcinoma tissues was significantly higher than that in the adjacent tissues (χ2=54.738, P<0.001). Moreover, the higher expression level of HIBADH protein was related to the larger tumor diameter, the higher tumor lymphatic invasion rate, the later pT stage, the higher the lymph node metastasis rate, and the later pTNM stage (P<0.05). HIBADH protein was also highly expressed in lymph nodes with metastatic carcinoma, and positive rate was 100% (48/48). The 10-year survival rate of patients in the HIBADH protein positive group and HIBADH protein negative group were 16.4% and 69.4%, respectively, which show the latter had a longer survival time (χ2=19.612, P<0.001). The migration capacity, invasion capacity, and scratch-wound capacity of the MKN45 cells were significantly decreased after HIBADH protein knockdown (P<0.05), but the proliferation capacity of the cells was not significantly changed (P>0.05). Conclusions The overexpression of HIBADH protein in gastric cancer suggests later tumor stage and poor prognosis. Inhibition expression of HIBADH protein can reduce the motility capacity of gastric cancer cells.

关键词: 3-羟异丁酸脱氢酶; 胃癌; 预后; 细胞运动

Key words: 3-hydroxyisobutyrate dehydrogenase; gastric cancer; prognosis; cell movement

登录后 ,请手动点击刷新查看图表内容。 没有账号,
1. Chen W, Zheng R, Baade PD, et al. Cancer statistics in China, 2015. CA Cancer J Clin, 2016, 66(2): 115-132.
2. Matsuoka T, Yashiro M. Biomarkers of gastric cancer: current topics and future perspective. World J Gastroenterol, 2018, 24(26): 2818-2832.
3. Magalhães H, Fontes-Sousa M, Machado M. Immunotherapy in advanced gastric cancer: an overview of the emerging strategies. Can J Gastroenterol Hepatol, 2018, 2018: 2732408.
4. Chao HC, Chung CL, Pan HA, et al. Protein tyrosine phosphatase non-receptor type 14 is a novel sperm-motility biomarker. J Assist Reprod Genet, 2011, 28(9): 851-861.
5. Rougraff PM, Paxton R, Kuntz MJ, et al. Purification and characterization of 3-hydroxyisobutyrate dehydrogenase from rabbit liver. J Biol Chem, 1988, 263(1): 327-331.
6. Rougraff PM, Zhang B, Kuntz MJ, et al. Cloning and sequence analysis of a cDNA for 3-hydroxyisobutyrate dehydrogenase. J Biol Chem, 1989, 264(10): 5899-5903.
7. Zhao C, Huo R, Wang FQ, et al. Identification of several proteins involved in regulation of sperm motility by proteomic analysis. Fertil Steril, 2007, 87(2): 436-438.
8. Martínez-Heredia J, de Mateo S, Vidal-Taboada JM, et al. Identification of proteomic differences in asthenozoospermic sperm samples. Hum Reprod, 2008, 23(4): 783-791.
9. Yue Q, Zhen H, Huang M, et al. Proteasome inhibition contributed to the cytotoxicity of arenobufagin after its binding with Na, K-ATPase in human cervical carcinoma HeLa cells. PLoS One, 2016, 11(7): e0159034.
10. Smyth EC, Verheij M, Allum W, et al. Gastric cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol, 2016, 27(suppl 5): v38-v49.
11. Chen DH, Yu JW, Wu JG, et al. Significances of contactin-1 expression in human gastric cancer and knockdown of contactin-1 expression inhibits invasion and metastasis of MKN45 gastric cancer cells. J Cancer Res Clin Oncol, 2015, 141(12): 2109-2120.
12. Du Y, Jiang B, Song S, et al. Metadherin regulates actin cytoskeletal remodeling and enhances human gastric cancer metastasis via epithelial-mesenchymal transition. Int J Oncol, 2017, 51(1): 63-74.
13. Song S, Pei G, Du Y, et al. Interaction between CD133 and PI3K-p85 promotes chemoresistance in gastric cancer cells. Am J Transl Res, 2018, 10(1): 304-314.
14. Pei G, Luo M, Ni X, et al. Autophagy facilitates metadherin-induced chemotherapy resistance through the AMPK/ATG5 pathway in gastric cancer. Cell Physiol Biochem, 2018, 46(2): 847-859.
15. Njau RK, Herndon CA, Hawes JW. New developments in our understanding of the beta-hydroxyacid dehydrogenases. Chem Biol Interact, 2001, 130-132(1-3): 785-791.
16. Li L, Dworkowski FS, Cook PF. Importance in catalysis of the 6-phosphate-binding site of 6-phosphogluconate in sheep liver 6-phosphogluconate dehydrogenase. J Biol Chem, 2006, 281(35): 25568-25576.
17. Chen YY, Ko TP, Chen WH, et al. Conformational changes associated with cofactor/substrate binding of 6-phosphogluconate dehydrogenase from Escherichia coli and Klebsiella pneumoniae: implications for enzyme mechanism. J Struct Biol, 2010, 169(1): 25-35.
18. Reitz S, Alhapel A, Essen LO, et al. Structural and kinetic properties of a beta-hydroxyacid dehydrogenase involved in nicotinate fermentation. J Mol Biol, 2008, 382(3): 802-811.
19. Tchigvintsev A, Singer A, Brown G, et al. Biochemical and structural studies of uncharacterized protein PA0743 from Pseudomonas aeruginosa revealed NAD+-dependent L-serine dehydrogenase. J Biol Chem, 2012, 287(3): 1874-1883.
20. Zhang Y, Zheng Y, Qin L, et al. Structural characterization of a β-hydroxyacid dehydrogenase from Geobacter sulfurreducens and Geobacter metallireducens with succinic semialdehyde reductase activity. Biochimie, 2014, 104: 61-69.
21. Murín R, Schaer A, Kowtharapu BS, et al. Expression of 3-hydroxyisobutyrate dehydrogenase in cultured neural cells. J Neurochem, 2008, 105(4): 1176-1186.
22. Tasi YC, Chao HC, Chung CL, et al. Characterization of 3-hydroxyisobutyrate dehydrogenase, HIBADH, as a sperm-motility marker. J Assist Reprod Genet, 2013, 30(4): 505-512.
23. Ishak Gabra MB, Yang Y, Lowman XH, et al. IKKβ activates p53 to promote cancer cell adaptation to glutamine deprivation. Oncogenesis, 2018, 7(11): 93.
24. Jiang Z, Zhang C, Gan L, et al. iTRAQ based quantitative proteomics approach identifies novel diagnostic biomarkers that were essential for glutamine metabolism and redox homeostasis for gastric cancer. Proteomics Clin Appl, 2018, [Epub ahead of print].
25. Xiao S, Zhou L. Gastric cancer: Metabolic and metabolomics perspectives (review). Int J Oncol, 2017, 51(1): 5-17.