miR­30c reduces myocardial ischemia/reperfusion injury by targeting SOX9 and suppressing pyroptosis.

MicroRNAs (miRNAs or miRs) are commonly involved in regulating myocardial ischemia/reperfusion (I/R) injury by binding and silencing their target genes. However, whether miRNAs regulate myocardial I/R-induced pyroptosis remains unclear. The present study established an in vivo rat model of myocardial I/R injury and in vitro hypoxia/reoxygenation (H/R) injury model in rat primary cardiomyocytes to investigate the function and the underlying mechanisms of miRNAs on I/R injury-induced pyroptosis. RNA sequencing was utilized to select the candidate miRNAs between normal and I/R group. Reverse transcription-quantitative PCR and western blotting were performed to detect candidate miRNAs (miR-30c-5p, also known as miR-30c) and SRY-related high mobility group-box gene 9 (SOX9) expression, as well as expression of pyroptosis-associated proteins (NF-κB, ASC, caspase-1, NLRP3) in the myocardial I/R model. ELISA was used to measure pyroptosis-associated inflammatory markers IL-18 and IL-1ß. Moreover, the link between miR-30c and SOX9 was predicted using bioinformatics and luciferase reporter assay. In myocardial I/R injured rats, miR-30c was downregulated, while the expression of SOX9 was upregulated. Overexpression of miR-30c inhibited pyroptosis both in vivo and in vitro. Furthermore, miR-30c negatively regulated SOX9 expression by binding its 3'untranslated region. In conclusion, the miR-30c/SOX9 axis decreased myocardial I/R injury by suppressing pyroptosis, which may be a potential therapeutic target.

Extracellular vesicles derived from human umbilical cord mesenchymal stem cells relieves diabetic retinopathy through a microRNA-30c-5p-dependent mechanism.

AIMS: Extracellular vesicle (EV)-transferred microRNAs (miRNAs) are proved to be potentially therapeutic candidates. Here, we attempted to unveil the role of delivery of miR-30c-5p by human umbilical cord mesenchymal stem cells (hUCMSCs)-derived EVs in diabetic retinopathy (DR). METHODS: miR-30c-5p and PLCG1 expression in streptozotocin-induced diabetes mellitus (DM) rats and high glucose (HG)-treated human retinal endothelial cells (HRECs) was quantified, followed by analysis on their interaction. EVs were isolated from hUCMSCs and co-cultured with HRECs. Through gain- and loss-of-function assays, the role of hUCMSCs-derived EV containing miR-30c-5p in DR involving PLCG1 and NF-κB pathway was analyzed in vitro and in vivo. RESULTS: Elevated PLCG1 was found in DM rats and HG-treated HRECs where miR-30c-5p was reduced while increased in hUCMSC-derived EVs. PLCG1 was pinpointed as a target gene of miR-30c-5p, which consequently disrupted the PKC/NF-κB pathway. hUCMSC-derived EVs decreased inflammation reaction by transferring miR-30c-5p in DM rats and HG-treated HRECs. Furthermore, similar changing tendency was observed in HG-treated HRECs induced by overexpressed miR-30c-5p through downregulation of PLCG1 in vivo. CONCLUSION: Overall, our findings underlined delivery of miR-30c-5p by hUCMSC-derived EVs as a novel suppressor in the inflammatory response following DR.

TGF­ß1 affects the renal cancer miRNome and regulates tumor cells proliferation.

TGF­ß1 is a pleiotropic cytokine that can either promote or inhibit cancer development and progression. It was previously found that TGF­ß1 can regulate the expression of several microRNAs (miR or miRNA) involved in the progression of renal cell carcinoma (RCC). Therefore, the present study aimed to analyze the effects of TGF­ß1 on the global RCC miRNome. It was found that TGF­ß1 can regulate a complex network consisting of miRNAs and mRNAs involved in RCC transformation. In particular, TGF­ß1 was revealed to regulate the proliferation of RCC cells while concomitantly modifying the expression of oncogenic regulators, including avian erythroblastosis virus E26 (V­Ets) oncogene homolog­1 (ETS1). In addition, TGF­ß1 was demonstrated to regulate the expression of a number of miRNAs including miR­30c­5p, miR­155­5p, miR­181a­5p and miR­181b­5p. By contrast, TGF­ß1 reciprocally modified the expression of genes encoding TGF­ß1 receptors and SMADs, indicating a novel regulatory feedback mechanism mediated through the miRNAs. These data suggested that ETS1 served different roles in different subtypes of RCC tumors, specifically by functioning as an oncogene in clear cell RCC while as a tumor suppressor in papillary RCC.

Effect of miR-30c/YWHAZ axis on drug resistance of pancreatic cancer cells / 中华内分泌外科杂志

Objective:To investigate whether microRNA-30c (miR-30c) mediates the resistance of pancreatic cancer cells to gemcitabine (Gb) by targeting tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activating protein zeta polypeptide (YWHAZ) .Methods:SW1990 cell line with the lowest expression of miR-30c in human pancreatic cancer cell lines was screened by RT-qPCR. Gb-resistant cell line SW1990/Gb was established and divided into SW1990/Gb group (untransfected) , miR-30c over expression (Ad-miR-30c) group, Ad-miR-30c negative control (Ad-eGFP) group, and SW1990 group. The level of miR-30c was measured by RT-qPCR; the half inhibitory concentration (IC50) and drug resistance index (IR) were measured by CCK-8 method; the expression of drug resistance-related protein P-gp, apoptosis-related protein Caspase-1, migration and transfer-related proteins MMP-9, YWHAZ and downstream pathway-kinase mitogen-activated protein kinase (p38MAPK) /extracellular regulatory protein kinase 1 (ERK1) protein was measured by Western blot. After co-transfection of Ad-miR-30c and YWHAZ overexpressing adenovirus (Ad-YWHAZ) , the expression of P-gp and YWHAZ pathway related proteins was measured by Western blot method.Results:The IC50 (59.16±5.14, nmol/L) , IR (11.15±0.19) , expressions of YWHAZ protein (1.59±0.15) and P-gp (2.43±0.26) in SW1990/Gb-resistant cells were high, the expression of miR-30c (0.25 ±0.02) was low ( P<0.05) , and the p38MAPK/ERK pathway was activated. After up-regulating the expression of miR-30c (1.59±0.15) in SW1990/Gb cells, the IC50 (25.14±2.15, nmol/L) and IR (5.48±0.12) , YWHAZ (1.49±0.13) , P-gp (1.46± 0.10) decreased ( P<0.05) , and the p38MAPK/ERK pathway was activated. Up-regulating the expression of YWHAZ could reverse the above-mentioned effects of Ad-miR-30c ( P<0.05) . Conclusions:The expression of miR-30c is low in pancreatic cancer Gb-resistant cell lines. Up-regulating the expression of miR-30c can target and inhibit the YWHAZ/p38MAPK/ERK pathway, inhibit the expression of drug-resistant protein P-gp, and reduce the resistance of pancreatic cancer cells to Gb.

Sevoflurane protects against cerebral ischemia/reperfusion injury via microrna-30c-5p modulating homeodomain-interacting protein kinase 1.

Sevoflurane (SEV) has been reported to be an effective neuroprotective agent for cerebral ischemia/reperfusion injury (CIRI). However, the precise molecular mechanisms of Sev preconditioning in CIRI remain largely unknown. Therefore, CIRI model was established via middle cerebral artery occlusion method. SEV was applied before modeling. after successful modeling, lentivirus was injected into the lateral ventricle of the brain. Neurological impairment score was performed in each group, and histopathologic condition, infarct volume, apoptosis, inflammation, oxidative stress, microRNA (miR)-30 c-5p and homeodomain-interacting protein kinase 1 (HIPK1) were detected. Mouse hippocampal neuronal cell line HT22 cells were pretreated with SEV, and the in vitro model was stimulated via oxygen-glucose deprivation and reoxygenation. The corresponding plasmids were transfected, and the cell growth was detected, including inflammation and oxidative stress, etc. The targeting of miR-30 c-5p with HIPK1 was examined. The results clarified that reduced miR-30 c-5p and elevated HIPK1 were manifested in CIRI. SEV could improve CIRI and modulate the miR-30 c-5p-HIPK1 axis in vitro and in vivo, and miR-30 c-5p could target HIPK1. Depressed miR-30 c-5p could eliminate the protection of SEV in vitro and in vivo. Repression of HIPK1 reversed the effect of reduced miR-30 c-5p on CIRI. Therefore, it is concluded that SEV is available to depress CIRI via targeting HIPK1 through upregulated miR-30 c-5p.

The RNA-binding protein hnRNPU regulates the sorting of microRNA-30c-5p into large extracellular vesicles.

The transfer of microRNAs (miRs) via extracellular vesicles (EVs) is a functionally relevant mechanism of intercellular communication that regulates both organ homoeostasis and disease development. Little is known about the packaging of miRs into EVs. Previous studies have shown that certain miRs are exported by RNA-binding proteins into small EVs, while for other miRs and for large EVs, in general, the export mechanisms remain unclear. Therefore, a proteomic analysis of endothelial cell-derived large EVs was performed, which revealed that heterogeneous nuclear ribonucleoprotein U (hnRNPU) is abundantly present in EVs. EVs were characterized by electron microscopy, immunoblotting and nanoparticle tracking analysis. Taqman microRNA array and single qPCR experiments identified specific miR patterns to be exported into EVs in an hnRNPU-dependent way. The specific role of hnRNPU for vesicular miR-sorting was confirmed independently by gain- and loss-of-function experiments. In our study, miR-30c-5p was the miR whose export was most significantly regulated by hnRNPU. Mechanistically, in silico binding analysis showed that the export of miRs into EVs depends on the binding efficiency of the respective miRs to hnRNPU. Among the exported miRs, a significant enrichment of the sequence motif AAMRUGCU was detected as a potential sorting signal. Experimentally, binding of miR-30c-5p to hnRNPU was confirmed independently by RNA-immunoprecipitation, electrophoretic mobility shift assay and reciprocally by miR-pulldown. Nuclear binding of miR-30c-5p to hnRNPU and subsequent stabilization was associated with a lower cytoplasmatic abundance and consequently reduced availability for vesicular export. hnRNPU-dependent miR-30c-5p export reduced cellular migration as well as pro-angiogenic gene expression in EV-recipient cells. In summary, hnRNPU retains miR-30c-5p and other miRs and thereby prevents their export into large EVs. The data presented provide a novel and functionally relevant mechanism of vesicular miR export.

Downregulation of microRNA-30c-5p was responsible for cell migration and tumor metastasis via COTL1-mediated microfilament arrangement in breast cancer.

BACKGROUND: Breast cancer metastasis is the main problem that affects the therapy and prognosis of breast cancer patients. Studies have indicated the role of microRNAs in breast cancer regulation, but the mechanisms are largely unknown. METHODS: In this study, we determined the expression of microRNA-30c-5p (miR-30c-5p) and coactosin-like protein 1 (COTL1) gene in breast cancer tissues, and revealed their effects on breast cancer metastasis regulation. Breast cancer and paracancerous tissues were collected. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to analyze the expression of miR-30c-5p and COTL1, and breast cancer cell line (MCF-7) was employed to verify the relationship between miR-30c-5p and COTL1. Western blot analysis and immunofluorescence were used for proteins analysis and microfilament observation, respectively. A dual-luciferase reporter gene was used for microRNA-gene interaction assay. RESULTS: The results showed that the expression of miR-30c-5p decreased, while the expression of COTL1 increased in breast cancer tissues. The results of luciferase reporting gene assay showed that, COTL1 was the target of miR-30c-5p. After miR-30c-5p was upregulated, the expression of COTL1 was reduced, microfilament arrangement was in disorder, and cell migration ability was inhibited. After miR-30c-5p was downregulated, the expression of COTL1 was increased, and the cell migration ability was enhanced. COTL1 protein expression levels were significantly higher in cancer tissues with lymph node metastasis. CONCLUSIONS: These findings indicate that miR-30c-5p/COTL1 pathway regulates breast cancer metastasis and can be used as a potential therapy target.

E2F7, regulated by miR­30c, inhibits apoptosis and promotes cell cycle of prostate cancer cells.

Prostate cancer (PCa) remains a leading cause of mortality among men in the United States and Western Europe. The molecular mechanism of PCa pathogenesis has not been fully elucidated. In the present study, the expression profile of E2F transcription factor 7 (E2F7) in PCa was examined using immunohistochemistry and reverse transcription­quantitative PCR, whilst cell cycle progression and apoptosis were determined using fluorescent cell activated sorting techniques. Cell viability was measured using Cell Counting Kit­8 in loss­ and gain­of­function studies. Dual­luciferase reporter assay was used to verify if E2F7 was one of the potential targets of miR­30c. The staining score of E2F7 of PCa tissues was found to be notably higher compared with that of adjacent normal tissues. Suppression of E2F7 expression in PCa cell lines led to significantly reduced proliferation rates, increased proportion of cells in the G1 phase of the cell cycle and higher apoptotic rates compared with those in negative control groups. Dual­luciferase reporter assay revealed E2F7 to be one of the binding targets of microRNA (miR)­30c. In addition, transfection of miR­30c mimics into PCa cells resulted in reduced cell viability, increased proportion of cells in the G1 phase and higher apoptotic rates. By contrast, transfection with the miR­30c inhibitor led to lower apoptosis rates of PCa cells compared with negative control groups, whilst E2F7 siRNA co­transfection reversed stimulatory effects of miR­30c inhibitors on cell viability. In addition, the expression of cyclin­dependent kinase inhibitor p21 were found to be upregulated by transfection with either E2F7 siRNA or miR­30c mimics into PCa cells. In conclusion, the present study suggested that E2F7 may be positively associated with PCa cell proliferation by inhibiting p21, whereas E2F7 is in turn under regulation by miR­30c. These observations suggest the miR­30c/E2F7/p21 axis to be a viable therapeutic target for PCa.

Role of MicroRNA-30c in cancer progression.

MicroRNAs (miRNAs or miRs) is a non-coding small RNA of a type of 18~24 nucleotide-regulated gene that has been discovered in recent years. It mainly degrades the target gene mRNA or inhibits its translation process through the complete or incomplete bindings with 3'UTR of target genes, followed by the regulation of individual development, apoptosis, proliferation, differentiation and other life activities through the post-transcriptional regulation. Among many miRNAs, the microRNA family, miR-30, plays diverse roles in these key process of neoplastic transformation, metastasis, and clinical outcomes in different cancer progression. As key member of miR-30, miR-30c is regulated by oncogenic transcription factors and cancer progression related genes. Recently, numerous studies have demonstrated that the aberrant expression of miR-30c was significantly associated with the majority of human cancer progression. In this review, the diverse roles of miR-30c in different cancer progression such as the cellular and molecular mechanisms, the potential applications in clinics were summarized to speculate the benefits of miR-30c over-expression in cancer treatment and prognosis.

LINC01342 promotes the progression of ovarian cancer by absorbing microRNA-30c-2-3p to upregulate HIF3A.

Ovarian cancer (OC) is a highly prevalent gynecologic malignancy and its mortality is extremely high. Therefore, the development of novel therapeutic approaches for OC is of great significance. In this study, LINC01342 was upregulated in OC tissue in the GSE38666 microarray and in tumor tissue samples collected in our center. The silencing of LINC01342 suppressed the proliferative and metastatic capacities of A2780 and HO8910 cells. Subcellular distribution assays showed that LINC01342 was mainly enriched in the cytoplasm. Subsequently, the downregulation of microRNA-30c-2-3p was proven to be the target of LINC01342. The silencing of microRNA-30c-2-3p enhanced the clonality and migratory capacity of OC cells. Moreover, the silencing of microRNA-30c-2-3p could reverse the inhibited migration and clonality in OC cells caused by LINC01342 knockdown. In addition, hypoxia-inducible factor 3 subunit α (HIF3A) was proven to be the target gene of microRNA-30c-2-3p, which was upregulated. HIF3A was negatively regulated by microRNA-30c-2-3p but positively regulated by LINC01342 in OC cells. An RNA binding protein immunoprecipitation assay showed that microRNA-30c-2-3p, LINC01342, and HIF3A could bind to argonaute RISC catalytic component 2. The overexpression of HIF3A reversed the inhibited migration and clonality in OC cells with LINC01342 knockdown. By analyzing the follow-up data from the enrolled OC patients, the LINC01342 and HIF3A levels were negatively correlated with prognosis, while the microRNA-30c-2-3p level was positively correlated with the same. In short, the upregulated LINC01342 in OC absorbs microRNA-30c-2-3p to release HIF3A. Thus, upregulated HIF3A expression accelerates the progression of OC.

MicroRNA-30c inhibits pancreatic cancer cell proliferation by targeting twinfilin 1 and indicates a poor prognosis.

BACKGROUND: Studies have reported that microRNA-30c (miR-30c) has vital functions in the development and progression of multiple cancers. AIM: To investigate the clinical significance and role of miR-30c in pancreatic cancer. METHODS: MiR-30c and twinfilin 1 (TWF1) expression levels were analyzed in Gene Expression Omnibus datasets and validated in human pancreatic cancer by quantitative real-time polymerase chain reaction (RT-qPCR). The effects of miR-30c on pancreatic cancer cell growth, apoptosis, and cell cycle were evaluated by CCK-8 and flow cytometry assays. Furthermore, the in vivo effects were investigated using a subcutaneous xenograft experiment. Target gene prediction software and luciferase reporter assays were used to identify TWF1 as a direct target of miR-30c. RESULTS: The expression of miR-30c was significantly decreased in pancreatic cancer tissues and associated with survival. Gain- and loss-of-function assays showed that miR-30c suppressed pancreatic cancer cell proliferation in vitro and in vivo. RT-qPCR, Western blot, and luciferase reporter assays showed that miR-30c directly targeted TWF1. The expression level of miR-30c was negatively correlated with TWF1 expression in pancreatic cancer tissues. Furthermore, the effects of ectopic miR-30c were rescued by TWF1 overexpression. CONCLUSION: Our results identified the role of the miR-30c/TWF1 axis in pancreatic cancer progression and demonstrated that miR-30c might serve as a prognostic biomarker and therapeutic target for pancreatic cancer.

MicroRNA-21 and microRNA-30c as diagnostic biomarkers for prostate cancer: a meta-analysis.

BACKGROUND: Prostate cancer (PCa) is the second leading cause of cancer death in men. Several articles have reported that microRNA-21 (miR-21) and microRNA-30c (miR-30c) have diagnostic values for PCa, but the results are inconclusive. In order to precisely assess the diagnostic values of miR-21 and miR-30c for PCa, this meta-analysis is performed. METHODS: Articles were searched in the databases of PubMed, Embase, and Web of Knowledge (search date: September 6, 2018). Studies were included if they were designed to evaluate the diagnostic performance of miR-21 or miR-30c for PCa. Using Stata 12.0 and Meta-Disc 1.4, the pooled sensitivity (SEN), specificity (SPE), positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR), and area under curve (AUC) of the summary receiver-operating characteristic (SROC) curve with the corresponding 95% CI were calculated. RESULTS: Overall, ten studies (six studies for miR-21 and four for miR-30c) involving1,371 participants were included in this meta-analysis. For miR-21, the pooled SEN and SPE were, respectively, 0.91 (95% CI: 0.87-0.94) and 0.88 (95% CI: 0.82-0.93), the pooled PLR and NLR were, respectively, 7.74 (95% CI: 4.81-12.47), 0.1 (95% CI: 0.06-0.15), the DOR was 77.64 (95% CI: 34.64-174.02), AUC of SROC was 0.95 (95% CI: 0.93-0.97). For miR-30c, the pooled SEN and SPE were, respectively, 0.74 (95% CI: 0.65-0.81) and 0.78 (95% CI: 0.72-0.83), the pooled PLR and NLR were, respectively, 3.39 (95% CI: 2.69-4.26), and 0.34 (95% CI: 0.26-0.44), the DOR was 10.06 (95% CI: 6.96-14.55), and AUC of SROC was 0.83 (95% CI: 0.79-0.86). CONCLUSION: For PCa, miR-21 is a good diagnostic biomarker and miR-30c is a moderate diagnostic biomarker.

Expression of miR-30c and BCL-9 in gastric carcinoma tissues and their function in the development of gastric cancer.

microRNA-30c (miR-30c) is a member of the miR-30s family, which is known to serve important roles in the occurrence and development of numerous tumor types. Our previous microarray analysis of extracted RNA from tissue samples was conducted to examine the expression of miR-30c and predict miR-30c target genes. In the present study, it was determined that the expression of miR-30c was differentially expressed in 82 paired gastric cancer (GC) and paracancerous tissues. Cellular expression of miR-30c in two GC cell lines MKN-45, MKN-74 and one non-cancer cell line GES-1 was modified using the miR-30c-mimic and miR-30c-inhibitor reagents, in a series of transfection experiments. Following transfection of cancer and non-cancer cell lines with the miR-30c-mimic, cell proliferation and apoptosis rates were increased. Compared with the NC group, MKN-74 cell proliferation was significantly inhibited (P<0.05) following transfection with the miR-30c-mimic at 48 and 24 h, GES-1 was significantly inhibited (P<0.05) at 24 and 48 h, and apoptosis was significantly reduced in transfected MKN-74 cells (P<0.05). The clinicopathological data and the expression of BCL-9 and miR-30c in patients with GC were used to identify associations. The expression levels of miR-30c were associated with age. Western blot analysis demonstrated that the BCL-9 expression levels in MKN-74 cells were higher following transfection with the miR-30c-mimic, and were lower following transfection with the miR-30c-inhibitor, both compared with the negative control group. It was concluded that compared with the negative control group, the expression of miR-30c was low in GC tissues and may be involved in GC development via regulation of proliferation, apoptosis and the cell cycle.

Mechanism of miR-30c over-expression inhibiting malignant phenotypes of cervical cancer cells / 中国病理生理杂志

AIM:To investigate the molecule mechanism of microRNA(miR)-30c over-expression inhibiting malignant phenotypes of cervical cancer cells.METHODS:Cervical cancer cell lines C33A,HeLa,SiHa and CaSki were transfected with pGenesil-1-miR-30c plasmid using Lipofectamine 2000 kit,and the expression of miR-30c was determined by TaqMan real-time PCR.The cell viability inhibition rate,colony formation ability,migration rate and apoptotic rate were measured by MTT assay,colony formation assay,Transwell experiment,and flow cytometry with Annexin V-FITC staining. The protein expression of Bax,Bcl-2, matrix metalloproteinase(MMP)-9, MMP-13 and tissue inhibitor of metalloprotei-nase-1(TIMP-1)was detected by Western blot.RESULTS:The expression levels of miRNA-30c in the cervical cancer cell lines transfected with pGenesil-1-miR-30c plasmid were significantly higher than those in negative control groups(cell lines transfected with pGenesil-1 plasmid)(P<0.01).Significantly increased cell viability inhibition rate,and decreased colony formation ability and migration rate were found in the cervical cancer cell lines over -expressing miR-30c as compared with negative control groups(P<0.05).The apoptotic rate in the cervical cancer cell lines over-expressing miR-30c was dramatically higher than that in control groups(P<0.05).Over-expression of miR-30c in cervical cancer cells promoted the protein expression of Bax and TIMP-1,and decreased the protein expression of Bcl-2 and MMP-13(P<0.05 or P<0.01).CONCLUSION:Over-expression of miR-30c significantly inhibits the viability and migration,and induces apop-tosis of cervical cancer cells.The mechanism may be related to activating apoptosis pathway and inhibiting MMP-13 protein expression.

Low expression of microRNA-30c promotes prostate cancer cells invasion involved in downregulation of KRAS protein.

Aberrant microRNA expression is associated with tumor development. The present study aimed to elucidate the role of miR-30c in the development of prostate cancer. Quantitative polymerase chain reaction was performed to compare miR-30c expression in LNCaP, DU145, PC-3 and RWPE-1 cell lines. Lentivirus expressing miR-30c was used to create stable overexpression cell lines to investigate the effects of miR-30c overexpression on cell proliferation, migration and invasion, which were determined in the prostate cancer cell line PC-3 by MTT, colony formation, wound healing and Transwell assays. Effects of miR-30c on KRAS were examined by western blot analysis. miR-30c expression was significantly lower (P<0.05) in the PC-3 cell line compared with LNCaP, DU145 and RWPE-1 cell lines. miR-30c overexpression in PC-3 inhibited tumor cell proliferation, migration and invasion in vitro. Furthermore, KRAS protein expression was downregulated in miR-30c overexpression cell lines compared with the negative control (NC) group (P<0.05). The present results demonstrated that overexpression of miR-30c inhibits prostate cancer cell line proliferation, migration and invasion, which was possibly caused by downregulation of KRAS protein by miR-30c. The data implicate miR-30c in the prognosis and treatment of prostate cancer.

Curcumin increases the sensitivity of Paclitaxel-resistant NSCLC cells to Paclitaxel through microRNA-30c-mediated MTA1 reduction.

Non-small-cell lung cancer is one of the most lethal cancers in the worldwide. Although Paclitaxel-based combinational therapies have long been used as a standard treatment in aggressive non-small-cell lung cancers, Paclitaxel resistance emerges as a major clinical problem. It has been demonstrated that Curcumin from Curcuma longa as a traditional Chinese medicine can inhibit cancer cell proliferation. However, the role of Curcumin in Paclitaxel-resistant non-small-cell lung cancer cells is not clear. In this study, we investigated the effect of Curcumin on the Paclitaxel-resistant non-small-cell lung cancer cells and found that Curcumin treatment markedly increased the sensitivity of Paclitaxel-resistant non-small-cell lung cancer cells to Paclitaxel. Mechanically, the study revealed that Curcumin could reduce the expression of metastasis-associated gene 1 (MTA1) gene through upregulation of microRNA-30c in Paclitaxel-resistant non-small-cell lung cancer cells. During the course, MTA1 reduction sensitized Paclitaxel-resistant non-small-cell lung cancer cells and enhanced the effect of Paclitaxel. Taken together, our studies indicate that Curcumin increases the sensitivity of Paclitaxel-resistant non-small-cell lung cancer cells to Paclitaxel through microRNA-30c-mediated MTA1 reduction. Curcumin might be a potential adjuvant for non-small-cell lung cancer patients during Paclitaxel treatment.

Serum microRNA-30c levels are correlated with disease progression in Xinjiang Uygur patients with chronic hepatitis B

We aimed to investigate the potential role and mechanism of microRNA-30c (miR-30c) in the pathological development of chronic hepatitis B (CHB). The serum levels of miR-30c in hepatitis B virus (HBV) carrier Xinjiang Uygur patients with inactive, low-replicative, high-replicative and HBe antigen-positive CHB were investigated. HepG2 cells were co-transfected with pHBV1.3 and miR-30c mimic or inhibitor or scramble RNA. The effects of miR-30c dysregulation on HBV replication and gene expression, cell proliferation and cell cycle were then investigated. miR-30c was down-regulated in Xinjiang Uygur patients with CHB compared to healthy controls and its expression level discriminated HBV carrier patients with inactive, low-replicative, high-replicative and HBe antigen-positive risk for disease progression. Overexpression of miR-30c significantly inhibited HBV replication and the expressions of HBV pgRNA, capsid-associated virus DNA and Hbx in hepatoma cells. Moreover, overexpression of miR-30c significantly inhibited cell proliferation and delayed G1/S phase transition in hepatoma cells. Opposite effects were obtained after suppression of miR-30c. Our results indicate that miR-30c was down-regulated in Xinjiang Uygur patients with CHB, and miR-30c levels could serve as a marker for risk stratification of HBV infection. Down-regulation of miR-30c may result in the progression of CHB via promoting HBV replication and cell proliferation.

MicroRNA-30c promotes natural killer cell cytotoxicity via up-regulating the expression level of NKG2D.

AIMS: Natural killer (NK) cells play critical roles in antitumor immunity. Our previous study showed that over-expression of miR-30c-1* enhanced NKL cell cytotoxicity through up-regulation of tumor necrosis factor-α via directly targeting transcription factor homeobox containing 1. MiR-30c, the complimentary microRNA of miR-30c-1*, has been found to exert regulatory effect on T cell function. However, the effect of miR-30c on NK cells is unknown. Therefore, this study aimed to investigate whether miR-30c could play a role to enhance NK cell activation and cytotoxicity. MAIN METHODS: Chemosynthesis exogenous miR-30c mimics and miR-30c inhibitor were transfected into NKL cells and isolated human peripheral blood NK cells, respectively. The expression levels of NK group 2, member D (NKG2D), CD107a and FasL on cell surface and cytotoxic ability of miRNAs transfected NKL cells against SMMC-7721 cells were evaluated. KEY FINDINGS: MiR-30c could increase the expression of NKG2D and CD107a on NKL cells, and enhance cytotoxic ability of NKL cells to kill SMMC-7721 cells. Moreover, miR-30c could up-regulate the expression of FasL on both NKL cells and human peripheral blood NK cells. However, the peripheral blood NK cells from only four in ten healthy donors appeared high expression levels of NKG2D and CD107a after miR-30c transfection. SIGNIFICANCE: MiR-30c could promote the cytotoxicity of NKL cells in vitro by up-regulating the expression levels of NKG2D, CD107a and FasL. However, the effect of miR-30c on ex vivo NK cells from different human individuals is diverse, indicating that miR-30c may play complicate and fine adjustment in immune system.

BCL9, a coactivator for Wnt/ß-catenin transcription, is targeted by miR-30c and is associated with prostate cancer progression.

B-cell lymphoma 9 (BCL9), a component of aberrantly activated Wnt signaling, is an important contributing factor to tumor progression. Our previous data indicated that downregulation of the tumor suppressor microRNA-30c (miR-30c) was a frequent pathogenetic event in prostate cancer (PCa). However, a functional link between miR-30c and BCL9/Wnt signaling, and their clinical and pathological significance in PCa, have not been well established. The present study demonstrated that miR-30c serves as a key negative regulator targeting BCL9 transcription in PCa cells. Ectopic expression of miR-30c was associated with reduced expression of Wnt pathway downstream targets, including c-Myc, cluster of differentiation 44 and sex determining region Y-box 9 in DU145 human PCa cells. Examination of clinical prostate specimens revealed higher levels of BCL9 expression in PCa compared with that in benign prostate tissues. After substantiating this finding by patient sample analysis, BCL9 expression or activity was observed to be closely correlated with PCa biochemical recurrence (BCR) and disease progression, whereas it was inversely associated with miR-30c. Furthermore, overexpression of BCL9 in PCa acted cooperatively with miR-30c low expression to predict earlier BCR in PCa. These findings indicate that inhibition of BCL9/Wnt signaling by miR-30c is important in the progression of PCa. Furthermore, the combined analysis of miR-30c and BCL9 may be valuable tool for prediction of BCR in PCa patients following radical prostatectomy.

Effects of miR-30 c on viability and migratory ability of HUVECs by tar-geting PAI-1 / 中国病理生理杂志

AIM: To investigate the effect of microRNA (miR)-30c on the viability and migratory ability of human umbilical vein endothelial cells (HUVECs) by targeting plasminogen activator inhibitor-1 (PAI-1).METHODS:The HUVECs were transfected with miR-30c mimic and inhibitor or negative control (NC), and then the expression levels of miR-30c, PAI-1 mRNA and protein were detected by RT-qPCR and Western blot.The viability and migratory ability of HUVECs were measured by CCK-8 assay and wound healing test .After bioinformatic analysis, the assessment of miR-30c binding to PAI-1 3’-UTR was carried out using a luciferase reporter gene assay .RESULTS:miR-30c directly down-regu-lated PAI-1 levels by binding to the 3’ UTR seed sequence of PAI-1 mRNA.Furthermore, transfection of a miR-30c mimic down-regulated the expression of PAI-1 at mRNA and protein levels, leading to enhanced migratory ability and viability of the HUVECs.However, transfection of a miR-30c inhibitor up-regulated the expression of PAI-1 at mRNA and protein le-vels, leading to decreased migratory ability and viability .CONCLUSION:Regulation of miR-30c level changes the migra-tory ability and viability of HUVECs by affecting the PAI-1 expression, indicating the involvement of miR-30c in modulating endothelial function .