HANR promotes lymphangiogenesis of hepatocellular carcinoma via secreting miR-296 exosome and regulating EAG1/VEGFA signaling in HDLEC cells.

The long noncoding RNA HANR has been shown to be involved in the progression of hepatocellular carcinoma (HCC). However, the underlying mechanism of HCC-associated long noncoding RNA (HANR)-regulated HCC metastasis and lymphangiogenesis has not been elucidated. RT-qPCR and Western blot methods were utilized to detect the gene expressions. Interaction of HANR with miR-296 was predicted by a bioinformatic program and validated by a dual-luciferase reporter assay. For the functional experiment, a transwell invasion assay was utilized to examine the invasion abilities of HepG2 and Huh-7 cells. The lymphatic vessel formation assay was used to show the HCC-associated lymphatic vessel formation ability of human dermal lymphatic endothelial cells (HDLEC). HANR was shown to directly bind to miR-296, and miR-296 downregulated HANR expression in HepG2 cells. Then, we observed that miR-296 inhibitor transfection in shHANR HCC cells could promote lymphatic vessel formation and invasion of HDLEC cells compared with shHANR HCC cells. EAG1 or VEGFA overexpression in HDLEC cells rescued lymphatic vessel formation and invasion in HDLEC cells coincubated with the medium of HepG2 cells expressing shHANR or miR-296 mimic. Ultimately, HANR knockdown and miR-296 mimic led to a significant decrease in the EAG1 and VEGFA expression levels in HepG2 cells. Here, we reveal a novel molecular mechanism in which the HANR/miR-296/EAG1/VEGF axis is responsible for the lymphangiogenesis of HCC cells. Our findings provide more insights into developing therapeutical or diagnostic methods by targeting HANR.

Shi Y ，Yang X ，Xue X ，Sun D ，Cai P ，Song Q ，Zhang B ，Qin L ... -  《-》

HANR promotes hepatocellular carcinoma progression via miR-214/EZH2/TGF-β axis.

LncRNA has been shown to associates with the initiation and progression of hepatocellular carcinoma (HCC). Recently, some studies showed that HANR function as an oncogene in HCC; however, the detailed mechanism of HANR-regulated HCC tumorigenesis and progression needs to be elucidated. We used RT-qPCR method to probe genes expression. MTT assay, wound healing assay and transwell invasion assay were utilized to examine proliferation and migration and invasion abilities of HepG2 cells. Xenograft tumor experiment was used to show the growth of tumors in vivo. HANR was evidently upregulated in HCC tumors and cells compared to normal tissues and cells. Besides, HANR knockdown induces attenuated cell proliferation, migration, invasion of HCC cells. By bioinformatic analysis and dual luciferase reporter assay, we found that miR-214 was the downstream target of HANR. Furthermore, miR-214 inhibitor largely enhanced tumor phenotypes of HCC cells regulated by HANR knockdown. HANR and miR-214 regulated the EZH2, then affecting TGFBR2 level. Finally, we demonstrated that EZH2 overexpression could greatly rescue HANR knockdown or miR-214 mimic-induced HCC tumorigenesis and progression. In this study, we report a newly identified regulatory mechanism HANR/miR-214/EZH2/TGF-β axis, which is implicated in tumorigenesis and progression of HCC. Our findings suggest that HANR facilitates the development of therapeutical strategies or diagnostic markers by targeting HANR.

Shi Y ，Yang X ，Xue X ，Sun D ，Cai P ，Song Q ，Zhang B ，Qin L ... -  《-》

LncRNA MCM3AP-AS1 Regulates Epidermal Growth Factor Receptor and Autophagy to Promote Hepatocellular Carcinoma Metastasis by Interacting with miR-455.

Zhang H ，Luo C ，Zhang G 《-》

Hsa_circ_0023404 enhances cervical cancer metastasis and chemoresistance through VEGFA and autophagy signaling by sponging miR-5047.

Cervical cancer has been shown to be one of the leading cancer-related death causes all over the world. Several studies demonstrates that hsa_circ_0023404 plays a crucial role in progression of cervical cancer; however, the detailed mechanism of hsa_circ_0023404 regulating cervical cancer metastasis and chemoresistance remains unclear. We used RT-qPCR and westernblot approach to detect expression levels of various genes in cervical tumors and cancer cells. To examine invasion and lymphatic vessel formation of Human Dermal Lymphatic Endothelial Cells (HDLEC), transwell invasion assay and lymphatic vessel assay were utilized in the presence of conditioned medium of HeLa and SiHa cells. To examine direct interaction between hsa_circ_0023404 and miR-5047, bioinformatic analysis and luciferase reporter assay were used. Besides, MTT and flow cytometry analysis were conducted to assess cell viability and apoptosis rate of HeLa cell. hsa_circ_0023404 knockdown attenuates invasion of cervical cancer cells and lymphatic vessel formation of HDLEC cells. hsa_circ_0023404 directly interacted with miR-5047. Moreover, miR-5047 inhibitor-transfected HeLa and SiHa cells enhanced invasion and lymphatic vessel formation of HDLEC cells. More interestingly, we confirmed that hsa_circ_0023404 knockdown and miR-5047 mimic downregulated the expression levels of VEGFA. The functional rescue experiments indicated VEGFA acted as key factor for hsa_circ_0023404- and miR-5047-regulated invasion and lymphatic vessel formaion. Ultimately, hsa_circ_0023404 and VEGFA were upregulated and showed positive correlation in cervical tumors, while miR-5047 was downregulated and showed negative correlation with hsa_circ_0023404 and VEGFA. On the other hand, autophagy-associated genes (Beclin1 and p62) were dysregulated in hsa_circ_0023404 depleted and overexpressed HeLa cells. hsa_circ_0023404 knockdown inhibited cell viability of cells, which was obviously abolished by autophagy inhibitor 3-MA in the presence of various concentrations of Cisplatin. Consistently, apoptosis rate was remarkably elevated in hsa_circ_0023404 depleted cells and diminished in hsa_circ_0023404 overexpressed cells under treatment of 2 μg/ml Cisplatin. Here, we reveal a novel role of hsa_circ_0023404 for cervical cancer metastasis and chemoresistance by regulating miR-5047. Our findings help understand mechanism underlying cervical cancer and development of therapeutical approaches for treating cervical cancer.

Guo J ，Chen M ，Ai G ，Mao W ，Li H ，Zhou J ... -  《-》

The Long Non-Coding RNA CASC2 Suppresses Cell Viability, Migration, and Invasion in Hepatocellular Carcinoma Cells by Directly Downregulating miR-183.

Hepatocellular carcinoma (HCC) is the most common malignant tumor of liver cells. Researchers have reported that cancer susceptibility candidate 2 (CASC2), a long non-coding RNA, is down-regulated in various cancers, including HCC. Our study aimed to investigate the molecular mechanism(s) of CASC2 in HCC. Real-time quantitative PCR (RT-qPCR) was used to analyze the expression of CASC2 and miR-183 in HCC tissues and cells. The viability of HCC SMMC-7721 and Huh-7 cells was detected through MTT assay. Colony formation assay was performed to assess the colony formation ability of HCC cells. The migration and invasion abilities of HCC cells were evaluated by Transwell assay. Western blot was conducted to examine levels of key Wnt/β-catenin signaling pathway factors, C-myc, cyclinD, survivin, and β-catenin. The interaction between CASC2 and miR-183 was affirmed by bioinformatics analysis and luciferase reporter assay. CASC2 was down-regulated in HCC tissues and cell lines, while miR-183 was up-regulated. The expression of miR-183 was negatively correlated with CASC2 expression in HCC tissues. Overexpression of CASC2 inhibited cell viability, colony formation, migration, and invasion in HCC cells, as well as Wnt/β-catenin signaling pathway activity. miR-183 was a downstream target of CASC2 and negatively regulated by CASC2. Introduction of miR-183 rescued CASC2-induced suppressive effects on HCC cell viability, colony formation, migration, and invasion and Wnt/β-catenin signaling. CASC2 inhibited cell viability and the colony formation, migration, and invasion abilities of HCC cells by directly downregulating miR-183 through inactivation of the Wnt/β-catenin signaling pathway.

Sun J ，Liu L ，Zou H ，Yu W ... -  《-》