Zheng Li, Chen Jiangtao, Zhou Zhongyong, He Zhikuan
Department of General Surgery, Huaihe Hospital of Henan University, Kaifeng, China.
Tumour Biol. 2017 May;39(5):1010428317705335. doi: 10.1177/1010428317705335.
Long non-coding RNA HOXD-AS1 (HOXD cluster antisense RNA 1) has been demonstrated to be closely associated with the progression of several tumors. However, the biological function of HOXD-AS1 and the underlying molecular mechanism in gastric cancer are still unclear. The expression of HOXD-AS1 in gastric cancer cell lines was evaluated by quantitative real-time polymerase chain reaction. The association of HOXD-AS1 expression and clinical parameters was statistically analyzed by chi-square test. Cell viability, colony formation capacity, and phosphorylation of Janus kinase 2 and signal transducer and activator of transcription 3 in treated SGC-7901 and BGC-823 cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, colony formation assay, and western blot analysis, respectively. The results indicated that HOXD-AS1 was significantly upregulated in gastric cancer cells and clinically involved in tumor size, invasion depth, tumor-node-metastasis stages, regional lymph nodes, lymphatic metastasis, as well as distant metastasis. HOXD-AS1 knockdown dramatically inhibited gastric cancer cell proliferation, colony formation capacity, and phosphorylation of Janus kinase 2 and signal transducer and activator of transcription 3 in vitro. In addition, HOXD-AS1 overexpression significantly promoted gastric cancer cell proliferation and colony formation capacity, whereas both Janus kinase small interfering RNAs and Janus kinase 2 inhibitor AG490 overturned these effects. Furthermore, xenograft assays confirmed the biological function of HOXD-AS1 in vivo. Taken together, our data elucidate that knockdown of HOXD-AS1 dramatically suppresses gastric cancer cell growth by inactivating the Janus kinase 2/signal transducer and activator of transcription 3 pathway in vitro and in vivo, contributing to a better understanding of gastric cancer pathogenesis and providing a possible theoretical foundation for long non-coding RNA-directed diagnosis and therapy against this disease.
长链非编码RNA HOXD-AS1（HOXD基因簇反义RNA 1）已被证明与多种肿瘤的进展密切相关。然而，HOXD-AS1在胃癌中的生物学功能及其潜在分子机制仍不清楚。通过定量实时聚合酶链反应评估HOXD-AS1在胃癌细胞系中的表达。采用卡方检验对HOXD-AS1表达与临床参数的相关性进行统计学分析。分别通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法、集落形成试验和蛋白质印迹分析检测处理后的SGC-7901和BGC-823细胞的细胞活力、集落形成能力以及Janus激酶2和信号转导及转录激活因子3的磷酸化水平。结果表明，HOXD-AS1在胃癌细胞中显著上调，且在临床上与肿瘤大小、浸润深度、肿瘤-淋巴结-转移分期、区域淋巴结、淋巴转移以及远处转移有关。HOXD-AS1基因敲低显著抑制了体外胃癌细胞的增殖、集落形成能力以及Janus激酶2和信号转导及转录激活因子3的磷酸化水平。此外，HOXD-AS1过表达显著促进了胃癌细胞的增殖和集落形成能力，而Janus激酶小干扰RNA和Janus激酶2抑制剂AG490均逆转了这些作用。此外，异种移植试验证实了HOXD-AS1在体内的生物学功能。综上所述，我们的数据表明，HOXD-AS1基因敲低通过在体外和体内使Janus激酶2/信号转导及转录激活因子3通路失活，显著抑制胃癌细胞生长，有助于更好地理解胃癌发病机制，并为针对该疾病的长链非编码RNA导向诊断和治疗提供可能的理论基础。
