Fang Zhixiong, He Langqiu, Jia Hui, Huang Qiusheng, Chen Dan, Zhang Zhiwei
Department of Infectious Disease, XiangTan City Central Hospital, XiangTan, People's Republic of China.
Edong Healthcare City Hospital of Traditional Chinese Medicine, Inefections Disease Hospital, Huangshi, People's Republic of China.
Iran J Basic Med Sci. 2017 Feb;20(2):187-192. doi: 10.22038/ijbms.2017.8246.
To explore the correlation between expression patterns and functions of miR-383 and LDHA in hepatocellular cancer (HCC).
We detected the expression of miR-383 and LDHA in 30 HCC tissues and their matched adjacent normal tissues using qRT-PCR. Then we performed MTT assay, foci formation assay, transwell migration assay, glucose uptake assay and lactate production assay to explore the function of miR-383 in cell proliferation, invasion and glycolysis in HCC cell lines. Luciferase reporter assay was used to explore whether LDHA was a target gene of miR-383. Western blot and qRT-PCR were used to further confirm LDHA was targeted by miR-383. Then the above functional experiments were repeated to see whether the function of LDHA could be inhibited by miR-383.
The results of qRT-PCR showed that miR-383 was down-regulated in HCC tissues compared with their matched adjacent normal tissues. Functional experiments showed that overexpression of miR-383 significantly suppressed cell proliferation, invasion and glycolysis. Luciferase reporter assay showed LDHA was a target gene of miR-383 and expression of LDHA was inversely correlated with that of miR-383 in HCC. Besides, increased cell proliferation, invasion and glycolysis triggered by LDHA could be inhibited by overexpression of miR-383 in HCC cell lines.
Our study proved that miR-383 is down-regulated in HCC and acts as a tumor suppressor through targeting LDHA. Targeting the miR-383-LDHA axis might be a promising strategy in HCC treatment.
探讨miR - 383与乳酸脱氢酶A（LDHA）在肝细胞癌（HCC）中的表达模式与功能之间的相关性。
我们采用qRT - PCR检测了30例HCC组织及其配对的癌旁正常组织中miR - 383和LDHA的表达。然后我们进行了MTT法、集落形成试验、Transwell迁移试验、葡萄糖摄取试验和乳酸生成试验，以探究miR - 383在HCC细胞系中的细胞增殖、侵袭及糖酵解功能。利用荧光素酶报告基因检测法探究LDHA是否为miR - 383的靶基因。采用蛋白质免疫印迹法（Western blot）和qRT - PCR进一步证实LDHA是miR - 383的靶标。随后重复上述功能实验，观察miR - 383是否能抑制LDHA的功能。
qRT - PCR结果显示，与配对的癌旁正常组织相比，miR - 383在HCC组织中表达下调。功能实验表明，miR - 383过表达显著抑制细胞增殖、侵袭及糖酵解。荧光素酶报告基因检测显示LDHA是miR - 383的靶基因，且在HCC中LDHA的表达与miR - 383呈负相关。此外，miR - 383在HCC细胞系中的过表达可抑制由LDHA引发的细胞增殖、侵袭及糖酵解增加。
我们的研究证明miR - 383在HCC中表达下调，并通过靶向LDHA发挥肿瘤抑制作用。靶向miR - 383 - LDHA轴可能是HCC治疗的一种有前景的策略
