Zhang Liyong, Ding Fang, Cao Wenfeng, Liu Zhongmin, Liu Wei, Yu Zaicheng, Wu Yu, Li Wendong, Li Yanda, Liu Zhihua
National Laboratory of Molecular Oncology, Cancer Institute and Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Clin Cancer Res. 2006 Mar 1;12(5):1639-46. doi: 10.1158/1078-0432.CCR-05-1858.
Stomatin-like protein 2 (SLP-2) is a novel and unusual stomatin homologue of unknown functions. It has been implicated in interaction with erythrocyte cytoskeleton and presumably other integral membrane proteins, but not directly with the membrane bilayer. We show here the involvement of SLP-2 in human esophageal squamous cell carcinoma (ESCC), lung cancer, laryngeal cancer, and endometrial adenocarcinoma and the effects of SLP-2 on ESCC cells.
Previous work of cDNA microarray in our laboratory revealed that SLP-2 was significantly up-regulated in ESCC. The expression of SLP-2 was further evaluated in human ESCC, lung cancer, laryngeal cancer, and endometrial adenocarcinoma by semiquantitative reverse transcription-PCR, Western blot, and immunohistochemistry. Mutation detection of SLP-2 exons was done by PCR and automated sequencing. Antisense SLP-2 eukaryotic expression plasmids were constructed and transfected into human ESCC cell line KYSE450. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, clonogenecity assay, flow cytometry assay, nude mice tumorigenetic assay, and cell attachment assay were done to investigate the roles of SLP-2 gene.
All tumor types we tested showed overexpression of SLP-2 compared with their normal counterparts (P < or = 0.05). Moreover, immunohistochemistry analysis of mild dysplasia, severe dysplasia, and ESCC showed that overexpression of SLP-2 occurred in premalignant lesions. Mutation analysis indicated that no mutation was found in SLP-2 exons. KYSE450 cells transfected with antisense SLP-2 showed decreased cell growth, proliferation, tumorigenecity, and cell adhesion.
SLP-2 was first identified as a novel cancer-related gene overexpressed in human ESCC, lung cancer, laryngeal cancer, and endometrial adenocarcinoma. Decreased cell growth, cell adhesion, and tumorigenesis in the antisense transfectants revealed that SLP-2 may be important in tumorigenesis.
类stomatin蛋白2（SLP - 2）是一种功能未知的新型且独特的stomatin同源物。它被认为与红细胞细胞骨架以及可能的其他整合膜蛋白相互作用，但不直接与膜双层相互作用。我们在此展示了SLP - 2在人食管鳞状细胞癌（ESCC）、肺癌、喉癌和子宫内膜腺癌中的作用以及SLP - 2对ESCC细胞的影响。
我们实验室先前的cDNA微阵列研究表明，SLP - 2在ESCC中显著上调。通过半定量逆转录 - PCR、蛋白质印迹法和免疫组织化学进一步评估SLP - 2在人ESCC、肺癌、喉癌和子宫内膜腺癌中的表达。通过PCR和自动测序进行SLP - 2外显子的突变检测。构建反义SLP - 2真核表达质粒并转染到人ESCC细胞系KYSE450中。进行3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐检测、克隆形成试验、流式细胞术检测、裸鼠致瘤试验和细胞黏附试验以研究SLP - 2基因的作用。
与正常对应组织相比，我们检测的所有肿瘤类型均显示SLP - 2过表达（P≤0.05）。此外，对轻度发育异常、重度发育异常和ESCC的免疫组织化学分析表明，SLP - 2的过表达发生在癌前病变中。突变分析表明，SLP - 2外显子未发现突变。用反义SLP - 2转染的KYSE450细胞显示细胞生长、增殖、致瘤性和细胞黏附能力下降。
SLP - 2首次被鉴定为在人ESCC、肺癌、喉癌和子宫内膜腺癌中过表达的新型癌症相关基因。反义转染细胞中细胞生长、细胞黏附和肿瘤发生能力的下降表明SLP - 2可能在肿瘤发生中起重要作用。
