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 ... -  《-》

circMTO1 promotes tumorigenesis and chemoresistance of cervical cancer via regulating miR-6893.

Cervical cancer has been one of the most common cancer in women worldwide. However, the detailed mechanism underlying circMTO1-regulated cervical cancer remains unclear. RT-qPCR and westernblot were used to examine genes expression levels. Wound healing assay and transwell invasion assay were utilized to probe migration and invasion abilities of cervical cancer cells. In addition, cell viability and apoptosis were measured by MTT and TUNEL assay, respectively. Finally, we employed bioinformatic approach to analyze gene expression levels in clinical cervical cancer tissues. circMTO1 was shown to be greatly upregulated in cervical cancer cell lines and tumors. circMTO1 directly interacts with miR-6893. miR-6893 could restore circMTO1-regulated migration, invasion and chemoresistance of cervical cancer cells. Furthermore, we identified S100A1 as downstream effector for circMTO1/miR-6893-induced tumorigenesis of cervical cancer cells. Besides, westernblot analyses demonstrated that circMTO1 and miR-6893 inhibitor enhanced Beclin1 expression and downregulated p62 level. We found that autophagy inhibitor 3-MA could revert cell viability and apoptosis of HeLa regulated by circMTO1 and miR-6893 inhibitor. More importantly, the bioinformatic results showed the dysregulated expression patterns of circMTO1, miR-6893 and S100A1 in human clinical cervical cancer tissues. In summary, we reveal, for the first time, a novel mechanism of circMTO1/miR-6893 in tumorigenesis and chemoresistance of cervical cancer. Our findings support the notion of developing the new therapies and biomarkers by targeting circMTO1 for cervical cancer.

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

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 ... -  《-》

Circular RNA hsa_circ_0000511 Improves Epithelial Mesenchymal Transition of Cervical Cancer by Regulating hsa-mir-296-5p/HMGA1.

As the second largest gynecological cancer, cervical cancer has been widely reported in recent years in which circular RNA is involved in the disease process. We earlier found that the expression of hsa_circ_0000511 in cervical cancer cells increased significantly, but its role in the process of cervical cancer is not clear. The purpose of this study is to explore its possible mechanisms in cervical cancer. Quantitative reverse transcription polymerase chain reaction (qRT-PCR), cell counting kit-8 assay, Transwell test, cell transfection, RNA pull-down assay and dual-luciferase reporter assay, and Western blot analysis were used to detect the expression and distribution of hsa_circ_0000511 in SiHa and HeLa cells, the ability of invasion and proliferation, and the modulated relationships between hsa_circ_0000511 and hsa-mir-296-5p, hsa-mir-296-5p, and HMGA1. hsa_circ_0000511 had the highest expression in SiHa and HeLa cells, and the expression in the cytoplasm was significantly higher than that in the nucleus, and its expression was not affected by RNase R. When hsa_circ_0000511 was silenced, its expression in SiHa and HeLa cells was significantly decreased; the proliferation, invasion, and migration abilities of the two kinds of cells were significantly enhanced; and the protein expression of E-cadherin was significantly upregulated, while the protein expression of N-cadherin was significantly downregulated. The expression of hsa-mir-296-5p was lower in SiHa and HeLa cells; however, its expression was increased when hsa_circ_0000511 was inhibited and decreased when hsa_circ_0000511 was overexpressed, so did the ability of proliferation, invasion, and migration and the protein expression of E-cadherin. Interestingly, the protein expression of HMGA1 also changed in these two cells when hsa-mir-296-5p was inhibited or overexpressed. Our results indicate that the upregulated hsa_circ_0000511 can inhibit the proliferation, invasion, and migration of SiHa and HeLa cells by regulating hsa-mir-296-5p/HMGA1, suggesting that the hsa_circ_0000511/hsa-mir-296-5p/HMGA1 pathway may be a potential target for the treatment of cervical cancer.

Xie J ，Chen Q ，Zhou P ，Fan W ... -  《-》

Overexpression of circular RNA hsa_circ_0001038 promotes cervical cancer cell progression by acting as a ceRNA for miR-337-3p to regulate cyclin-M3 and metastasis-associated in colon cancer 1 expression.

Cervical cancer (CC) is a common cancer threatens women's health worldwide. Circular RNAs (circRNAs) is critically involved in carcinogenesis of various cancers. This work aimed to explore the expression pattern, functions and mechanisms of hsa_circ_0001038 in CC. RT-qPCR was performed to evaluate the levels of hsa_circ_0001038 in CC cell lines and tissues. Kaplan-Meier curves was applied to evaluate the overall survival rate of CC patients with high or low expression of hsa_circ_0001038. Fisher's exact test was analyzed to explore the relationship of hsa_circ_0001038 expression and CC patients' clinical features. Loss/Gain-of function assays were used to evaluate the role of hsa_circ_0001038 on the growth, apoptosis, migration and invasion of CC cell lines. The competing endogenous RNA (ceRNA) mechanism was predicted by bioinformatics databases and verified by dual-luciferase reporter assay. We found that hsa_circ_0001038 was highly expressed in CC cells and tissues and elevated hsa_circ_0001038 was closely related to the clinical severity including lymph node invasion and myometrial invasion. In addition, overexpressed hsa_circ_0001038 correlated with unfavorable outcome in CC patients. Knockdown of hsa_circ_0001038 attenuated cell growth, migration, and invasion but induced cell apoptosis. Ectopic expression of hsa_circ_0001038 increased cell oncogenic properties. For mechanism investigation, hsa_circ_0001038 could sponge miR-337-3p to release its suppression on cyclin-M3 (CNNM3) and metastasis-associated in colon cancer 1 (MACC1), thereby promoting CC cell growth and invasive potential, respectively. In conclusion, hsa_circ_0001038 plays an oncogenic role in CC cells partly by activating CNNM3 and MACC1.

Wang Y ，Wang L ，Wang W ，Guo X ... -  《-》