Wen Ruiling, Xiao Yingying, Zhang Yuhua, Yang Min, Lin Yongping, Tang Jun
KingMed Diagnostics and KingMed School of Laboratory Medicine, Guangzhou Medical University, Guangzhou, Guangdong 510330, P.R. China.
Cytate Institute for Precision Medicine and Innovation, Guangzhou Cytate Biomedical Technologies Inc., Guangzhou, Guangdong 510663, P.R. China.
Oncol Rep. 2016 Dec;36(6):3172-3180. doi: 10.3892/or.2016.5135. Epub 2016 Sep 28.
Tubulin tyrosine ligase like 12 (TTLL12), a member of the tubulin tyrosine ligase (TTLL) family, has not been completely characterized to date. It is reported that histone methylation, tubulin modifications, mitotic duration and chromosome ploidy play crucial roles in a variety of cancers, and are related to tumorigenesis and cancer progression. A recent study showed that TTLL12 may be a pseudo-enzyme which has a SET-like domain and a TTL-like domain. In the present study, we first used 3'-rapid amplification of cDNA ends (3'-RACE) to amplify the transcripts of the TTLL12 gene from a human lung cancer cell line H1299, and unexpectedly discovered a new transcript isoform characterized with an additional 108-bp nucleotide sequence inserted at the location from 902 to 903 bases of the TTLL12 coding sequence (CDS), where it also locates between exons 5 and 6. Next, utilizing RT-PCR and Sanger sequencing, we further confirmed the existence of such a new transcript isoform of TTLL12 in more human cancer cells including lung cancer cells and other cancer cells. Moreover, several lung cancer cell lines were found to display a much higher proportion of the new isoform compared with TTLL12 wild-type transcript. These results suggest that the new TTLL12 isoform may be of importance for proper maintenance of lung cancer cells. Therefore, the new isoform of TTLL12, with the inserted sequences probably acting as a disordered region, provides a novel perspective regarding TTLL12 functions in human cancers including lung cancer.
微管蛋白酪氨酸连接酶样蛋白12(TTLL12)是微管蛋白酪氨酸连接酶(TTLL)家族的成员,迄今为止尚未得到完全表征。据报道,组蛋白甲基化、微管蛋白修饰、有丝分裂持续时间和染色体倍性在多种癌症中起关键作用,并且与肿瘤发生和癌症进展相关。最近的一项研究表明,TTLL12可能是一种具有SET样结构域和TTL样结构域的假酶。在本研究中,我们首先使用cDNA末端快速扩增技术(3'-RACE)从人肺癌细胞系H1299中扩增TTLL12基因的转录本,意外地发现了一种新的转录本异构体,其特征是在TTLL12编码序列(CDS)的第902至903个碱基位置插入了一个额外的108个碱基的核苷酸序列,该位置也位于外显子5和6之间。接下来,利用逆转录聚合酶链反应(RT-PCR)和桑格测序,我们进一步证实了这种新转录本异构体在更多人类癌细胞(包括肺癌细胞和其他癌细胞)中的存在。此外,发现几种肺癌细胞系中这种新异构体的比例比TTLL12野生型转录本高得多。这些结果表明,新的TTLL12异构体可能对肺癌细胞的正常维持很重要。因此,TTLL12的新异构体,其插入序列可能作为一个无序区域,为TTLL12在包括肺癌在内的人类癌症中的功能提供了一个新视角。