Methyl ferulic acid ameliorates ethanol-induced L02 cell steatosis through microRNA-378b-mediated CaMKK2-AMPK pathway / 中国中药杂志

Alcoholic liver disease(ALD), with its increasing morbidity and mortality, has seriously and extensively affected the health of people worldwide. Methyl ferulic acid(MFA) has been proven to significantly inhibit alcohol-induced lipid production in L02 cells through the AMP-activated protein kinase(AMPK) pathway, but its in-depth mechanism remains unclear. This study aimed to further clarify the mechanism of MFA in improving lipid accumulation in L02 cells through the microRNA-378b(miR-378b)-mediated calcium/calmodulin-dependent protein kinase kinase 2(CaMKK2)-AMPK signaling pathway based on existing researches. L02 cells were induced by 100 mmol·L~(-1) ethanol for 48 h to establish the model of ALD in vitro, and 100, 50, and 25 μmol·L~(-1) concentration of MFA was treated. MiR-378b plasmids(containing the overexpression plasmid-miR-378b mimics, silence plasmid-miR-378b inhibitor, and their respective negative control-miR-378b NCs) were transfected into L02 cells by electroporation to up-regulate or down-regulate the levels of miR-378b in L02 cells. The levels of total cholesterol(TC) and triglyceride(TG) in cells were detected by commercial diagnostic kits and automatic biochemical analyzers. The expression levels of miR-378b in L02 cells were detected by real-time quantitative polymerase chain reaction(qRT-PCR). CaMKK2 mRNA levels were detected by PCR, and protein expressions of related factors involved in lipid synthesis, decomposition, and transport in lipid metabolism were detected by Western blot. The results displayed that ethanol significantly increased TG and TC levels in L02 cells, while MFA decreased TG and TC levels. Ethanol up-regulated the miR-378b level, while MFA effectively inhibited the miR-378b level. The overexpression of miR-378b led to lipid accumulation in ethanol-induced L02 cells, while the silence of miR-378b improved the lipid deposition induced by ethanol. MFA activated the CaMKK2-AMPK signaling pathway by lowering miR-378b, thus improving lipid synthesis, decomposition, and transport, which improved lipid deposition in L02 cells. This study shows that MFA improves lipid deposition in L02 cells by regulating the CaMKK2-AMPK pathway through miR-378b.

Relationship between serum miR-199a-5p and miR-378a-3p levels and recurrence of infants with proliferative facial hemangioma after propranolol withdrawal / 口腔疾病防治

Objective@# To investigate the relationship between serum miR-199a-5p and miR-378a-3p levels and the recurrence of infants with proliferative facial hemangioma relapsed after propranolol withdrawal in infants.@*Methods@#Ninety-three infants with proliferative facial hemangioma were selected, all of whom received propranolol treatment. The recurrence of the hemangioma after treatment was followed up, and the children were divided into a recurrence group (23 patients) and a nonrecurrence group (70 patients). Venous blood was collected before and after treatment, and the serum levels of miR-199a-5p and miR-378a-3p were detected by qRT-PCR, and the relationship between the serum levels of miR-199a-5p and miR-378a-3p and the recurrence of proliferative facial hemangioma relapsed after propranolol withdrawal in infants was analyzed.@*Results @# The serum expressions levels of miR-199a-5p and miR-378a-3p in non-relapsed group were increased after treatment compared with before treatment (P < 0.05), and there was no significant difference in the levels of miR-199a-5p and miR-378a-3p in the serum of the recurrence group after treatment compared with before treatment (P > 0.05). After treatment, the levels of miR-199a-5p and miR-378a-3p in the serum of the recurrence group were lower than those of the nonrecurrence group (P < 0.05). After treatment, the serum levels of miR-199a-5p and miR-378a-3p in patients with Ⅲ ~Ⅳ Norm grade hemangioma were higher than those in patients with Ⅰ~Ⅱ Norm grade hemangioma (P < 0.05). Logistic regression analysis showed that the low expression of miR-199a-5p, low expression of miR-378a-3p and tumor grade Ⅰ~Ⅱ after treatment were risk factors for the recurrence of proliferative facial hemangioma after propranolol withdrawal in infants (P < 0.05).@*Conclusion@# Low serum levels of miR-199a-5p and miR-378a-3p are associated with the recurrence of proliferative facial hemangioma after propranolol withdrawal in infants.

MicroRNA-378a-3p Downregulation as a Novel Biomarker with Poor Clinical Outcomes in Cervical Cancer / 生物医学与环境科学（英文）

Objective@#Cervical cancer (CC) is one of the most common malignant tumors in gynecology. This study aimed to investigate the prognostic significance of serum microRNA (miR)-378a-3p in CC and the effect of miR-378a-3p on tumor growth.@*Methods@#Real-time quantitative polymerase chain reaction analysis was used to measure the expression of miR-378a-3p in serum from patients with CC and healthy control subjects as well as from CC tissues and adjacent normal tissues. The association between serum miR-378a-3p levels and clinicopathological factors was analyzed. The correlation between miR-378a-3p levels and overall survival (OS) of CC patients was determined by Kaplan-Meier analysis. The CC cell proliferation and migration abilities after transfection of miR-378a-3p mimics were detected by Cell Counting Kit-8 and scratch wound healing assays, respectively. Tumor volume and weight in mice treated with miR-378a-3p were measured using a caliper and an electronic balance.@*Results@#MiR-378a-3p expression was downregulated in the serum and tissues of CC patients compared to that in healthy control subjects and normal tissues, respectively. Low expression of miR-378a-3p was positively correlated with large tumor size, advanced tumor stage, and lymph node metastasis. The OS of patients with low expression of miR-378a-3p was significantly lower than that of patients with high expression. Overexpression of miR-378a-3p suppressed the proliferation and migration of CC cells. @*Conclusion@#MiR-378a-3p downregulation is associated with the development and prognosis of CC, suggesting that it may be a potential biomarker for CC.

miR-378a-5p and miR-630 induce lens epithelial cell apoptosis in cataract via suppression of E2F3

Cataract, an eye disease that threatens the health of millions of people, brings about severe economic burden for patients and society. MicroRNA (miR)-378a-5p and miR-630 were recognized as essential regulators in multiple cancers. However, the exact functions of miR-378a-5p and miR-630 in cataract are still unclear. The expression of miR-378a-5p, miR-630, and E2F transcription factor 3 (E2F3) in tissues and cells was measured by quantitative real-time polymerase chain reaction. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay was used to evaluate cell viability. Flow cytometry was conducted to analyze cell apoptosis. The interaction between E2F3 and miR-378a-5p or miR-630 was confirmed by dual-luciferase reporter assay. The expression of proteins E2F3, B cell lymphoma (Bcl-2), Bcl-2 associated X (Bax), and cleaved caspase 3 was detected by western blot assay. The expression of miR-378a-5p and miR-630 was up-regulated whereas E2F3 was down-regulated in human cataract lens tissues compared with normal lens tissues. Depletion of miR-378a-5p or miR-630 enhanced proliferation and reduced apoptosis of human lens epithelial cells. Interestingly, up-regulation of E2F3 exhibited the same trend. Next, dual-luciferase reporter assay validated the interaction between E2F3 and miR-378a-5p or miR-630. The rescue experiments further revealed that E2F3 knockdown could recover miR-378a-5p, and miR-630 inhibitor induced promotion of cell proliferation and inhibition of apoptosis in cataract. miR-378a-5p and miR-630 repressed proliferation and induced apoptosis of lens epithelial cells by targeting E2F3 in cataract, representing a prospective alternative therapy for cataract.

Clinical application value of combined detection of serum miR-378 and miR-21 in gastric cancer / 中华肿瘤杂志

Objective@#To investigate the clinical value of combined detection of serum miR-378 and miR-21 in gastric cancer (GC).@*Methods@#Eighty-seven patients with GC and 78 patients with colorectal cancer(CRC) from National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences were selected, 83 individuals undergoing healthy physical examination were selected as the healthy controls. The levels of serum miR-378 and miR-21 were detected by quantitative real-time PCR (RT-qPCR) (result data were transformed as log2 for analysis).@*Results@#Relative expression levels of miR-378 in the serum were -1.24, -3.25 and -2.73 in healthy controls, GC and CRC patients, respectively. Compared with the healthy controls, the levels of serum miR-378 were significantly decreased in GC and CRC patients (both P<0.05). Relative expression levels of miR-21 in the serum were 0.11, 2.34 and 2.47 in healthy controls, GC and CRC patients, respectively. Compared with the healthy controls, the levels of serum miR-21 were significantly up-regulated in GC and CRC patients (both P<0.05). Moreover, the serum level of miR-378 in GC patients was inversely associated with tumor clinical stage (P<0.05). However, the level of miR-21 showed no significant differences among patients with different clinical and pathological characteristics (all P>0.05). The area under the receiver operating characteristic curve (AUC), sensitivity and specificity of miRNA-378 to diagnose GC was 0.770, 82.0% and 66.0%, respectively, and were 0.900, 85.0%, and 88.0% of miR-21, respectively. The AUC, sensitivity and specificity of combined detection of serum miR-378 and miR-21 to diagnose GC were 0.930, 92.0% and 87.0%, respectively, while the AUC of combined detection of serum CEA and CA-199 was 0.767, the AUC of combined all of the four factors was 0.946.@*Conclusion@#The combined detection of serum miR-378 and miR-21 have a certain effect on diagnosis of GC.

microRNA-378 promotes proliferation and migration of renal carcinoma cells and its mechanism / 肿瘤

Objective: To investigate the effects of microRNA-378 (miR-378) on proliferation and migration of renal cancer cell lines in vitro, and to explore the possible mechanism. Methods: The expression of miR-378 in renal cell carcinoma and normal renal epithelial tissues was analyzed using the datasets from The Cancer Genome Atlas (TCGA). The expression of miR-378 in human renal carcinoma cell lines 786-O, ACHN, 769-P and Caki-1 as well as human renal proximal tubular epithelial cell line HK2 was detected by real-time fluorescent quantitative PCR (RFQ-PCR). MiR-378 mimic and inhibitor were synthesized and then transfected into renal carcinoma 786-O and Caki-1 cells, respectively. The proliferation and migration of 786-O and Caki-1 cells were detected by CCK-8 assay, Transwell chamber assay and wound healing assay, respectively. The potential target gene of miR-378 was screened by bioinformatics, and confirmed by RFQ-PCR and Western blotting in renal cancer cells, respectively. Results: The expression level of miR-378 in renal cancer tissues and cell lines was significantly up-regulated as compared with that in normal renal epithelial tissues or normal renal epithelial HK2 cells (all P < 0.001). After transfection with miR-378 mimic, the proliferation (P < 0.01) and migration (P < 0.001) of 786-O cells were remarkably increased. In contrast, after transfection with miR-378 inhibitors, the proliferation (P < 0.01) and migration (P < 0.001) of Caki-1 cells were remarkably suppressed. By bioinformatics assay, the large tumor suppressor 2 (LATS2) was identified as a target gene of miR-378 in renal cancer. The mRNA and protein expression of LATS2 were negatively related with the expression of miR-378 in renal cancer 786-O and Caki-1 cells (all P < 0.01). Conclusion: Overexpression of miR-378 may promote the proliferation and migration of renal cancer cells by negative targeting LATS2 gene.

Expression and Clinical Significance of miR-224 and miR-378e in Colorectal Cancer Tissues / 天津医药

Objective To investigate the expression and clinical significance of microRNA-224 and microRNA-378e in colorectal cancer tissues and normal mucosa adjacent to tumor lesions. Methods The gene chip technology was used to detect the different expression of miRNA in colorectal carcinoma tissues and adjacent normal tissues, which was then confirmed by real-time PCR. The relationship between the pathology and clinical data was analyzed. Results The expres-sion level of miR-224 was significantly up-regulated in tumor tissue, while miR-378e was down-regulated in tumor tissue, which was confirmed by real-time PCR. The expression of miR-224 was strongly associated with histological types, while miR-378e was strongly associated with the infiltration depth of colorectal cancer. Conclusion miR-224 is a potent tumor promoter, while miR-378e is a potent tumor suppressor. Both miR-224 and miR-378e can be used as potential colorectal cancer molecular markers.