MiR-21 Indicates Poor Prognosis in Tongue Squamous Cell Carcinomas as an Apoptosis Inhibitor
MiR-21 Indicates Poor Prognosis in Tongue Squamous Cell Carcinomas as an Apoptosis Inhibitor
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MiR-21 作为凋亡抑制剂表明舌鳞状细胞癌的预后不良。
DOI：
10.1158/1078-0432.ccr-08-3053
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发表时间：
2009-06-15
影响因子：
11.5
通讯作者：
Song, Erwei
中科院分区：
文献类型：
--
作者：
Li, Jinsong;Huang, Hongzhang;Song, Erwei
目的：我们的目的是研究miR-21在舌鳞状细胞癌（TSCC）的表达，并与患者的临床状态，并探讨其对TSCC细胞生长，凋亡和tumorogenicity.Experimental Design的贡献：microRNA分析在10例TSCC与微阵列。采用定量逆转录-PCR对103例患者的miR-21过表达进行定量，并与患者的病理临床状态相关。免疫组化检测TPM 1和PTEN的表达，末端脱氧核苷酸转移酶介导的dUTP标记检测细胞凋亡。此外，将miR-21反义寡核苷酸（阿索）转染到SCC-15和CAL 27细胞系中，并通过3-（4，5-二甲基噻唑-2基）-2，5-二苯基溴化四氮唑、贴壁集落形成和软琼脂测定来测定肿瘤细胞生长，而通过膜联蛋白V测定、细胞色素c释放和半胱天冬酶3测定来测定细胞凋亡。结果：miR-21在TSCC中的表达明显高于癌旁正常组织。miR-21的表达水平与TPM 1和PTEN的表达及癌细胞凋亡密切相关。多因素分析显示，miR-21表达是一个独立的预后因素，表明生存不良。在TSCC细胞系中用阿索抑制miR-21降低了存活率和锚定非依赖性生长，并诱导TSCC细胞系的凋亡。用siRNA同时沉默TPM 1仅部分重现了miR-21阿索的作用。结论：miR-21是TSCC的独立预后指标，可能通过沉默TPM 1抑制癌细胞凋亡而在TSCC的发生发展中发挥作用。
Purpose: We aim to examine miR-21 expression in tongue squamous cell carcinomas (TSCC) and correlate it with patient clinical status, and to investigate its contribution to TSCC cell growth, apoptosis, and tumorigenesis.Experimental Design: MicroRNA profiling was done in 10 cases of TSCC with micro-array. MiR-21 overexpression was quantitated with quantitative reverse transcription-PCR in 103 patients, and correlated to the pathoclinical status of the patients. Immunohistochemistry was used to examine the expression of TPM1 and PTEN, and terminal deoxynucleotidyl transferase-mediated dUTP labeling to evaluate apoptosis. Moreover, miR-21 antisense oligonucleotide (ASO) was transfected in SCC-15 and CAL27 cell lines, and tumor cell growth was determined by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide, adherent colony formation, and soft agar assay, whereas apoptosis was determined by Annexin V assay, cytochrome c release, and caspase 3 assay. Tumorigenesis was evaluated by xenografting SCC-15 cells in nude mice.Results: MiR-21 is overexpressed in TSCC relative to adjacent normal tissues. The level of miR-21 is reversely correlated with TPM1 and PTEN expression and apoptosis of cancer cells. Multivariate analysis showed that miR-21 expression is an independent prognostic factor indicating poor survival. Inhibiting miR-21 with ASO in TSCC cell lines reduces survival and anchorage-independent growth, and induces apoptosis in TSCC cell lines. Simultaneous silencing of TPM1 with siRNA only partially recapitulates the effect of miR-21 ASO. Furthermore, repeated injection of miR-21 ASO suppresses tumor formation in nude mice by reducing cell proliferation and inducing apoptosis.Conclusions: miR-21 is an independent prognostic indicator for TSCC, and may play a role in TSCC development by inhibiting cancer cell apoptosis partly via TPM1 silencing.