Identification of PTK6, via RNA Sequencing Analysis, as a Suppressor of Esophageal Squamous Cell Carcinoma
Identification of PTK6, via RNA Sequencing Analysis, as a Suppressor of Esophageal Squamous Cell Carcinoma
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DOI：
10.1053/j.gastro.2012.06.007
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发表时间：
2012-09-01
期刊：
影响因子：
29.4
通讯作者：
Guan, Xin-Yuan
中科院分区：
文献类型：
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作者：
Ma, Stephanie;Bao, Jessie Y. J.;Guan, Xin-Yuan
背景与目的：食管鳞状细胞癌（ESCC）是食管癌最常见的组织学亚型。ESCC被认为是通过许多遗传改变的积累而发展的，包括肿瘤抑制基因的失活和癌基因的激活。我们寻找与非肿瘤组织相比，在人ESCC样本中发生改变的转录本。方法：我们使用来自3名患者和邻近非肿瘤组织的ESCC样本进行整合转录组测序（RNA-Seq）分析，以鉴定ESCC组织中改变的转录本。我们进行了分子和功能研究的成绩单确定和调查的机制改变。研究结果：我们确定蛋白酪氨酸激酶6（PTK 6）作为一种转录本，在食管鳞癌组织和细胞系与非肿瘤组织或永生化正常食管细胞系相比，显着下调。PTK 6基因的启动子在食管鳞癌组织中至少部分通过超甲基化和组蛋白去乙酰化而失活。使用小发夹RNA敲低KYSE 30 ESCC细胞中的PTK 6增加了它们在培养物中形成病灶、迁移和侵入细胞外基质以及在裸鼠中形成肿瘤的能力。PTK 6在这些细胞中的过表达降低了它们在培养物中的增殖和小鼠中的肿瘤形成。PTK 6减少Akt和糖原合成酶激酶（GSK）3 β的磷酸化，导致β-连环蛋白的活化。结论：PTK 6被鉴定为通过PTK 6位点的表观遗传修饰在人ESCC组织中下调的转录物。其产物似乎通过减少Akt和GSK 3 β的磷酸化来调节细胞增殖，导致β-连环蛋白的活化。PTK 6水平降低促进小鼠异种移植肿瘤的生长;它可能被开发为ESCC的标志物。
BACKGROUND & AIMS: Esophageal squamous cell carcinoma (ESCC) is the most commonly observed histologic subtype of esophageal cancer. ESCC is believed to develop via accumulation of numerous genetic alterations, including inactivation of tumor suppressor genes and activation of oncogenes. We searched for transcripts that were altered in human ESCC samples compared with nontumor tissues. METHODS: We performed integrative transcriptome sequencing (RNA-Seq) analysis using ESCC samples from 3 patients and adjacent nontumor tissues to identify transcripts that were altered in ESCC tissue. We performed molecular and functional studies of the transcripts identified and investigated the mechanisms of alteration. RESULTS: We identified protein tyrosine kinase 6 (PTK6) as a transcript that was significantly down-regulated in ESCC tissues and cell lines compared with nontumor tissues or immortalized normal esophageal cell lines. The promoter of the PTK6 gene was inactivated in ESCC tissues at least in part via hypermethylation and histone deacetylation. Knockdown of PTK6 in KYSE30 ESCC cells using small hairpin RNAs increased their ability to form foci, migrate, and invade extracellular matrix in culture and form tumors in nude mice. Overexpression of PTK6 in these cells reduced their proliferation in culture and tumor formation in mice. PTK6 reduced phosphorylation of Akt and glycogen synthase kinase (GSK)3 beta, leading to activation of beta-catenin. CONCLUSIONS: PTK6 was identified as a transcript that is down-regulated in human ESCC tissues via epigenetic modification at the PTK6 locus. Its product appears to regulate cell proliferation by reducing phosphorylation of Akt and GSK3 beta, leading to activation of beta-catenin. Reduced levels of PTK6 promote growth of xenograft tumors in mice; it might be developed as a marker of ESCC.