microRNA-184 enhances the sensitivity of pheochromocytoma-12 cells to doxorubicin by targeting ADAM22

Zhao Nairui,Su Na,Wang Guangya,Fu Dongxia,Gao Fang,Zhang Yunna 대한독성 유전단백체 학회 2021 Molecular & cellular toxicology Vol.17 No.2

Background Pheochromocytoma (PCC) is a catecholamine-producing and neuroendocrine tumor with the 5-year overall survival of advanced stage PCC lower than 40%. Increasing evidence has shown that aberrant expression of microRNAs (miRNAs) plays important roles in the development and chemotherapy resistance of cancers including PCC. Objective The tumor-suppressive function of miR-184 has been identified in several types of cancers. The aim of this study is to explore the function and the underlying mechanism of miR-184 in the chemo-resistance of PCC. Results miR-184 expression was significantly lower in doxorubicin (Dox)-resistant pheochromocytoma-12 (PC-12) cells and PCC patients. Consistently, in vitro analysis showed that overexpression of miR-184 obviously improved the sensitivity of PC-12/Dox cells, while knockdown of miR-184 sensitised PC-12/Dox cells to chemotherapeutics. To further understand the possible functional mechanism of miR184 in the chemo-resistance of PCC, the targets of miR-184 were predicted. The results of miRDB database suggested A disintegrin and metalloproteinase 22 (ADAM22) carrying the potential complementary binding sites of miR-184 within its 3′-untranslated region (UTR). Further experiments confirmed that miR-184 bound the 3′-UTR of ADAM22 mRNA and down-regulated the expression of ADAM22 in PC-12/Dox cells. Moreover, ADAM22 was overexpressed in Dox-resistant PC-12 cells and PCC patients. Additionally, overexpression of ADAM22 attenuated miR-184-mediated chemo-sensitivity of PC-12/Dox cells. Conclusion miR-184 played a role in the chemo-sensitivity of PC-12/Dox cells at least partially via negatively regulating ADAM22. These results suggested miR-184 as a possible novel target to attenuate the chemo-resistance of PCC. Background Pheochromocytoma (PCC) is a catecholamine-producing and neuroendocrine tumor with the 5-year overall survival of advanced stage PCC lower than 40%. Increasing evidence has shown that aberrant expression of microRNAs (miRNAs) plays important roles in the development and chemotherapy resistance of cancers including PCC. Objective The tumor-suppressive function of miR-184 has been identified in several types of cancers. The aim of this study is to explore the function and the underlying mechanism of miR-184 in the chemo-resistance of PCC. Results miR-184 expression was significantly lower in doxorubicin (Dox)-resistant pheochromocytoma-12 (PC-12) cells and PCC patients. Consistently, in vitro analysis showed that overexpression of miR-184 obviously improved the sensitivity of PC-12/Dox cells, while knockdown of miR-184 sensitised PC-12/Dox cells to chemotherapeutics. To further understand the possible functional mechanism of miR184 in the chemo-resistance of PCC, the targets of miR-184 were predicted. The results of miRDB database suggested A disintegrin and metalloproteinase 22 (ADAM22) carrying the potential complementary binding sites of miR-184 within its 3′-untranslated region (UTR). Further experiments confirmed that miR-184 bound the 3′-UTR of ADAM22 mRNA and down-regulated the expression of ADAM22 in PC-12/Dox cells. Moreover, ADAM22 was overexpressed in Dox-resistant PC-12 cells and PCC patients. Additionally, overexpression of ADAM22 attenuated miR-184-mediated chemo-sensitivity of PC-12/Dox cells. Conclusion miR-184 played a role in the chemo-sensitivity of PC-12/Dox cells at least partially via negatively regulating ADAM22. These results suggested miR-184 as a possible novel target to attenuate the chemo-resistance of PCC.

MicroRNA-22 negatively regulates LPS-induced inflammatory responses by targeting HDAC6 in macrophages

윤기수,Jong Kook Park,Chae Yeon Lee,장재희,윤상호,Hyeok Yil Kwon,최수영,박진서 생화학분자생물학회 2020 BMB Reports Vol.53 No.4

Dysregulation of histone deacetylase 6 (HDAC6) can lead to the pathologic states and result in the development of various diseases including cancers and inflammatory diseases. The objective of this study was to elucidate the regulatory role of microRNA-22 (miR-22) in HDAC6-mediated expression of proinflammatory cytokines in lipopolysaccharide (LPS)-stimulated macrophages. LPS stimulation induced HDAC6 expression, but suppressed miR-22 expression in macrophages, suggesting possible correlation between HDAC6 and miR-22. Luciferase reporter assays revealed that 3’UTR of HDAC6 was a bona fide target site of miR-22. Transfection of miR-22 mimic significantly inhibited LPS-induced HDAC6 expression, while miR-22 inhibitor further increased LPS-induced HDAC6 expression. LPS-induced activation of NF-B and AP-1 was inhibited by miR-22 mimic, but further increased by miR-22 inhibitor. LPS-induced expression of pro-inflammatory cytokines such as TNF-, IL-1, and IL-6 was inhibited by miR-22 mimic, but further increased by miR-22 inhibitor. Taken together, these data provide evidence that miR-22 can downregulate LPS-induced expression of proinflammatory cytokines via suppression of NF-B and AP-1 axis by targeting HDAC6 in macrophages.

Overexpression of Long Non-Coding RNA MIR22HG Represses Proliferation and Enhances Apoptosis via miR-629-5p/TET3 Axis in Osteosarcoma Cells

( Haoliang Zhao ),( Ming Zhang ),( Xuejing Yang ),( Dong Song ) 한국미생물 · 생명공학회 2021 Journal of microbiology and biotechnology Vol.31 No.10

In this study, we evaluated the mechanism of long non-coding RNA MIR22 host gene (LncRNA MIR22HG) in osteosarcoma cells. Forty-eight paired osteosarcoma and adjacent tissues samples were collected and the bioinformatic analyses were performed. Target genes and potential binding sites of MIR22HG, microRNA (miR)-629-5p and tet methylcytosine dioxygenase 3 (TET3) were predicted by Starbase and TargetScan V7.2 and confirmed by dual-luciferase reporter assay. Cell Counting Kit-8, colony formation and flow cytometry assays were utilized to determine the viability, proliferation and apoptosis of transfected osteosarcoma cells. Pearson’s analysis was introduced for the correlation analysis between MIR22HG and miR-629-5p in osteosarcoma tissue. Relative expressions of MIR22HG, miR-629-5p and TET3 were measured by quantitative real-time polymerase chain reaction or Western blot. MiR-629-5p could competitively bind with and was negatively correlated with MIR22HG, the latter of which was evidenced by the high expression of miR-629-5p and low expression of MIR22HG in osteosarcoma tissues. Overexpressed MIR22HG repressed the viability and proliferation but enhanced apoptosis of osteosarcoma cells, which was reversed by miR-629-5p upregulation. TET3 was the target gene of miR-629-5p, and the promotive effects of upregulated miR-629-5p on the viability and proliferation as well as its repressive effect on apoptosis were abrogated via overexpressed TET3. To sum up, overexpressed MIR22HG inhibits the viability and proliferation of osteosarcoma cells, which was achieved via regulation of the miR-629- 5p/TET3 axis.

NUP210 and MicroRNA-22 Modulate Fas to Elicit HeLa Cell Cycle Arrest

Qiao Gu,Wenjie Hou,Huan Liu,Lijuan Shi,Zonghao Zhu,Wenfeng Ye,Xiaoyuan Ni 연세대학교의과대학 2020 Yonsei medical journal Vol.61 No.5

Purpose: Cervical cancer is one of the most fatal diseases among women in under-developed countries. To improve cervical cancer treatment, discovery of new targets is needed. In this study, we investigated the expression of NUP210, miR-22, and Fas in cervical cancer tissues and their functions in cell cycle regulation. Materials and Methods: We detected and compared the expression levels of NUP210, miR-22, and Fas in cervical cancer tissues with paired normal tissues using immunohistochemistry, Western blot, and real-time quantitative polymerase chain reaction. NUP210 was knocked down in HeLa cells via lentivirus, followed by cell cycle and proliferation analysis. Using a luciferase reporter assay, we explored the link between miR-22 and NUP210. We overexpressed miR-22 in HeLa cells and analyzed cell cycle and proliferation function. We then overexpressed miR-22 in NUP210 knockdown cells to explore the connection between Fas and miR-22-NUP210 signaling. Results: We found that NUP210 was overexpressed in cervical cancer patients. Knocking down NUP210 restored cell apoptosis and proliferation. We confirmed miR-22 as a regulator of NUP210 and verified that miR-22 was inhibited in cervical cancer development. We also found that restoring miR-22 expression could induce cell apoptosis. Finally, we found that miR-22-regulated expression of NUP210 could alter Fas expression and, in turn, elicit cell cycle arrest and proliferation. Conclusion: miR-22 in cervical cancer is downregulated, resulting in NUP210 overexpression and inhibition of Fas-induced cell apoptosis.

miR-215 Enhances HCV Replication by Targeting TRIM22 and Inactivating NF-κB Signaling

Hui Tian,Zhenkun He 연세대학교의과대학 2018 Yonsei medical journal Vol.59 No.4

Purpose: Hepatitis C virus (HCV) infection is a major cause of liver disease. Several miRNAs have been found to be associatedwith HCV infection. This study aimed to investigate the functional roles and possible molecular mechanisms of miR-215 in HCVreplication. Materials and Methods: The expression levels of miR-215 and TRIM22 were detected by quantitative real-time PCR (qRT-PCR)and western blot analysis in Con1b subgenomic genotype 1b HCV replicon cells (Con1b cells) and JFH1 full genome infectingHuh7.5.1 cells (Huh7.5.1 cells). HCV RNA levels were measured by qRT-PCR. The protein levels of NS3, NS5A, p65 subunit of NF-κB (p65), and phosphorylated p65 (p-p65) were determined by western blot analysis. The relationship between miR-215 andTRIM22 were explored by target prediction and luciferase reporter analysis. Results: miR-215 overexpression enhanced HCV replication in Con1b cells, while miR-215 knockdown suppressed HCV replicationin Huh7.5.1 cells. TRIM22 was confirmed to be a direct target of miR-215. TRIM22 upregulation resulted in a decline in HCVreplication, while TRIM22 inhibition led to enhancement of HCV replication. Additionally, exogenous expression of TRIM22 reversedthe facilitating effect of miR-215 on HCV replication, while TRIM22 downregulation counteracted the inhibitory effect ofmiR-215 knockdown on HCV replication. Furthermore, miR-215 targeted TRIM22 to block the NF-κB pathway, and exerted apositively regulatory role on HCV replication. Conclusion: miR-215 facilitated HCV replication via inactivation of the NF-κB pathway by inhibiting TRIM22, providing a novelpotential target for HCV infection.

Diagnostic Value of Circulating Extracellular miR-134, miR-185, and miR-22 Levels in Lung Adenocarcinoma-Associated Malignant Pleural Effusion

신윤미,윤지은,이옥준,한혜숙,임성남,안진영,이기형,이기만,최강현 대한암학회 2014 Cancer Research and Treatment Vol.46 No.2

PurposeThe accurate and timely diagnosis of malignant pleural effusion (MPE) in lung cancerpatients is important because MPE has a poor prognosis and is classified as stage IVdisease. Molecular biomarkers for pleural effusion, such as circulating extracellularmicroRNAs (miRNAs) isolated from pleural fluid, may help in the diagnosis of MPE. The present study examined whether miRNAs that are deregulated in lung cancer(miR-134, miR-185, and miR-22) can serve as diagnostic markers for lung adenocarcinoma-associated MPE (LA-MPE). Materials and MethodsReal-time reverse transcription quantitative polymerase chain reaction was used tomeasure the expression of the three miRNAs in samples from 87 patients with pleuraleffusion comprising 45 LA-MPEs and 42 benign pleural effusions (BPEs). The areaunder the receiver operating characteristic curve (AUC) was then used to evaluate thediagnostic performance of each of the three miRNAs and compare it with that of thecommon tumor marker, carcinoembryonic antigen (CEA). ResultsThe expression of all three miRNAs was significantly lower in LA-MPE than in BPE(p <0.001). The AUCs for miR-134, miR-185, miR-22, and CEA were 0.721, 0.882,0.832, and 0.898, respectively. Combining CEA with the three miRNAs increased thediagnostic performance, yielding an AUC of 0.942 (95% confidence interval, 0.864to 0.982), with a sensitivity of 91.9% and a specificity of 92.5%. ConclusionThe present study suggests that the expression levels of circulating extracellular miR-134, miR-185, and miR-22 in patients with pleural effusion may have diagnostic valuewhen differentiating between LA-MPE and BPE.

LncRNA-IMAT1 Promotes Invasion of Meningiomas by Suppressing KLF4/hsa-miR22-3p/Snai1 Pathway

Tao Zhang,Yu Ge,Daijun Wang,Qin Liu,Shuchen Sun,Lingyang Hua,Jiaojiao Deng,Shihai Luan,Haixia Cheng,Qing Xie,Ye Gong,Tao Zhang 한국분자세포생물학회 2022 Molecules and cells Vol.45 No.6

Malignant meningiomas often show invasive growth that makes complete tumor resection challenging, and they are more prone to recur after radical resection. Invasive meningioma associated transcript 1 (IMAT1) is a long noncoding RNA located on Homo sapiens chromosome 17 that was identified by our team based on absolute expression differences in invasive and non-invasive meningiomas. Our studies indicated that IMAT1 was highly expressed in invasive meningiomas compared with non-invasive meningiomas. In vitro studies showed that IMAT1 promoted meningioma cell invasion through the inactivation of the Krüppel-like factor 4 (KLF4)/hsa-miR22-3p/Snai1 pathway by acting as a sponge for hsa-miR22-3p, and IMAT1 knockdown effectively restored the tumor suppressive properties of KLF4 by preserving its tumor suppressor pathway. In vivo experiments confirmed that IMAT1 silencing could significantly inhibit the growth of subcutaneous tumors and prolong the survival period of tumor-bearing mice. Our findings demonstrated that the high expression of IMAT1 is the inherent reason for the loss of the tumor suppressive properties of KLF4 during meningioma progression. Therefore, we believe that IMAT1 may be a potential biological marker and treatment target for meningiomas.