Effects of miRNA-30a-5p and metadherin on proliferation, invasion and migration abilities of breast cancer cells / 肿瘤研究与临床

Objective:To investigate the effects of miRNA-30a-5p (miR-30a-5p) and metadherin (MTDH) on the proliferation, invasion and migration abilities of human breast cancer cells in vitro.Methods:The expression of MTDH in cancer and paracancerous tissues of 112 breast cancer patients in the database and miR-30a-5p in cancer and paracancerous tissues of 103 breast cancer patients in the database were analyzed using data from The Cancer Genome Atlas (TCGA) database. Pearson correlation analysis was used to analyze the correlation between miR-30a-5p and MTDH in 1 222 breast cancer patients in the database; the data were updated to August 2022. Breast cancer MDA-MB-231 cells were divided into negative control group (transfected with negative control sequence), miR-30a-5p overexpression group (transfected with miR-30a-5p mimics), siMTDH group [transfected with small interfering RNA against MTDH (siMTDH)], siMTDH+miR-30a-5p overexpression group (transfected with both siMTDH and miR-30a-5p mimics); cell proliferation ability was detected by methyl thiazol tetrazolium (MTT) assay, cell migration ability was detected by cell scratch assay, cell invasion ability was detected by Transwell assay. The relative expressions of miR-30a-5p, MTDH, matrix metalloproteinase (MMP)-2, MMP-9, vimentin and β-catenin mRNA in cells were detected by quantitative real-time fluorescence polymerase chain reaction (qRT-PCR), and the expressions of MTDH, N-cadherin (N-cad), β-catenin, Snail and MMP-9 proteins were detected by Western blotting.Results:In the TCGA database, MTDH expression level was higher and miR-30a-5p expression level was lower in breast cancer tissues compared with paracancerous tissues, and the differences were statistically significant (both P < 0.001). There was a negative correlation between MTDH and miR-30a-5p expressions in 1 222 patients with breast cancer ( r=-0.134, P < 0.001). Compared with the negative control group, the cell proliferation ability was reduced in both siMTDH group and miR-30a-5p overexpression group at 24, 48 and 72 h (all P < 0.001). The cell scratch healing rate in miR-30a-5p overexpression group and siMTDH group was lower than that in negative control group [(61.6±1.6)%, (54.7±5.9)% vs. (80.3±3.0)%] (both P < 0.05). Compared with the negative control group, The number of migrated cells in miR-30a-5p overexpression group and siMTDH group was less than that in negative control group (881±50, 725±63 vs. 1 172±66) (both P < 0.05). Compared with the negative control group, the relative expressions of MMP-2, MMP-9, vimentin and β-catenin mRNA were all down-regulated in MDA-MB-231 cells of miR-30a-5p overexpression group and siMTDH group (all P < 0.05). Compared with the negative control group, the relative expressions of N-cad, β-catenin, Snail and MMP-9 proteins were down-regulated in MDA-MB-231 cells of miR-30a-5p overexpression group and siMTDH group (all P < 0.05). There was no statistical difference in the number of migrated MDA-MB-231 cells between siMTDH+miR-30a-5p overexpression group and siMTDH group (476±5 vs. 389±46, t = 3.37, P = 0.078). There was no statistical difference in the number of migrated cells between siMTDH+miR-30a-5p overexpression group and miR-30a-5p overexpression group (476±5 vs. 477±22, t = 0.02, P = 0.983). Conclusions:The expression of miR-30a-5p is negatively correlated with the expression of MTDH in breast cancer tissues, and either overexpression of miR-30a-5p or silence of MTDH in breast cancer MDA-MB-231 cells in vitro can inhibit cell proliferation, invasion and migration, but MTDH may not be a target gene of miR-30a-5p.

Effect of telmisartan on expression of metadherin in the kidney of mice with unilateral ureter obstruction / 南方医科大学学报

OBJECTIVE@#To explore the effect of telmisartan on the expression of metadherin in the kidney of mice with unilateral ureter obstruction.@*METHODS@#Eighteen male C57 mice were randomized into sham-operated group, model group and telmisartan treatment group. In the latter two groups, renal interstitial fibrosis as the result of unilateral ureter obstruction (UUO) was induced by unilateral ureteral ligation with or without telmisartan intervention. Renal pathological changes of the mice were assessed using Masson staining, and immunohistochemistry and Western blotting were used to detect the expression of extracellular matrix proteins and metadherin in the kidney of the mice. In the experiment, cultured mouse renal tubular epithelial cells (mTECs) were stimulated with transforming growth factor-β1 (TGF-β1) and transfected with a siRNA targeting metadherin, and the changes in the expressions of extracellular matrix proteins and metadherin were detected using Western blotting.@*RESULTS@#The expressions of extracellular matrix proteins and metadherin increased significantly in the kidney of mice with UUO ( < 0.05). Intervention with telmisartan significantly lowered the expressions of extracellular matrix proteins and metadherin and alleviated the pathology of renal fibrosis in mice with UUO ( < 0.05). In cultured mTECs, siRNA-mediated knockdown of metadherin obviously reversed TGF-β1-induced increase in the expressions of extracellular matrix proteins and metadherin.@*CONCLUSIONS@#Telmisartan can suppress the production of extracellular matrix proteins and the expression of metadhein to attenuate UUO-induced renal fibrosis in mice.

Calcium channel blocker diltizem transiently inhibits migration and up-regulates metadherin expression in hepatocellular carcinoma cells / 南方医科大学学报

OBJECTIVE@#To investigate the effect of calcium channel blocker diltizem in reversing multi-drug resistance (MDR) and on metadherin expression in hepatocellular carcinoma cells and explore the molecular mechanism.@*METHODS@#Hepatocellular carcinoma MHCC97H and 7402 cells were treated with diltiazem hydrochloride, a calcium channel blocker (0, 25, 50, 100, 200, and 400 μmol/L), for 12, 24, or 48 h. Wound healing assay was employed to assess the changes in the mobility and migration of the cells following the treatments, and the changes in the expression levels of metadherin mRNA and protein and P-gp protein were determined using RT-PCR and immunocytochemistry.@*RESULTS@#Diltiazem hydrochloride could transiently inhibit the migration and movement of MHCC97H and 7402 cells in a time-and concentration-dependent manner ( < 0.05). Diltiazem hydrochloride at different concentrations also transiently up-regulated the expressions of metadherin mRNA and protein but did not inhibit the expression of P-gp protein in MHCC97H and 7402 cells.@*CONCLUSIONS@#Calcium channel blocker can transiently inhibit the migration of hepatocellular carcinoma cells and up-regulate the expression of metadherin mRNA and protein through a feedback mechanism, suggesting the potential risk of calcium channel blockers for promoting tumor progression during the treatment of malignant tumors.

The mechanisms of paeonol reversing multidrug resistance in ovarian cancer SKOV3/DDP cells / 药学学报

The purpose of this research is to investigate the effects and mechanisms of paeonol (PL), a phenolic compound found in many traditional Chinese formulations, on reversing drug resistance in the ovarian cancer resistant SKOV3/DDP cells. The results showed that PL had significant drug-resistant reversal effect on SKOV3/DDP cells. Flow cytometry showed that PL could inhibit P-glycoprotein (P-gp) function in a concentration-dependent manner. Fluorescent quantitative PCR and cell immunofluorescence techniques were used to detect mechanisms of action. Results revealed that both the inhibitory effect on MDR1/P-gp and metadherin (MTDH) expression and the induction effect on phosphatase and tensin homolog (PTEN), by 15, 30, and 60 μmol·L-1 PL, were increased with increased concentrations of PL (P P -1 PL treated group; however, the inhibition or induction effect on MTDH or PTEN protein were only comparable to the 15 μmol·L-1 PL treated group. The present study shows that the effect of PL on SKOV3/DDP cells may be related to the inhibition of P-gp function and expression, the inhibition of MDR1, MTDH expression, and the induction of PTEN expression, all which can provide a theoretical foundation for PL as a drug resistance reversal agent on the treatment of ovarian cancer chemotherapy resistance.

Expressions of metadherin and cyclinD1 in esophageal squamous cell carcinoma and their clinical significances / 肿瘤研究与临床

Objective To study the expressions of metadherin (MTDH) and cyclinD1 in esophageal squamous cell carcinoma (ESCC) and their clinical significances. Methods The protein expressions of MTDH and cyclinD1 were detected by immunohistochemistry in 78 cases of ESCC. Results The positive expression rate of MTDH in ESCC was 71.79%(56/78) and the positive expression rate of cyclinD1 in ESCC was 74.36%(58/78). The expressions of MTDH and cyclinD1 were significantly correlated with the degree of differentiation and lymph node metastasis (both P 0.05). Conclusion The over expressions of MTDH and cyclinD1 protein may involve in the occurrence and development of esophageal carcinoma, which play important roles in the invasion and metastasis of esophageal cancer.

MIR-375 modulates trastuzumab-resistance through EMT in HER2-positive breast cancer cells / 肿瘤

Objective: To investigate the role of microRNA-375 (miR-375) in the occurrence of trastuzumab-resistance in human epidermal growth factor receptor 2 (HER2)-positive breast cancer cells through regulating epithelial-mesenchymal transition (EMT). Methods: The sensitivities of HER2-positive breast cancer cell lines SK-BR-3 (a parental sensitive cell line) and SK-BR-3R (a trastuzumab-resistant cell line) to trastuzumab were detected by MTT assay. The expression levels of miR-375 and EMT-associated proteins E-cadherin and vimentin in the two cell lines were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. After miR-375 mimic was transfected into SK-BR-3R cells, the changes of miR-375, E-cadherin and vimentin expression levels were detected by real-time fluorescent quantitative PCR and Western blotting, and the change of trastuzumab sensitivity of SK-BR-3R cells was detected by MTT method. The potential target genes of miR-375 were predicted by bioinformatics software, and metadherin (MTDH) was selected as one of target genes. Luciferase reporter assay was conducted to verify the role of miR-375 in regulation of MTDH gene transcription. In addition, the changes of E-cadherin and vimentin expression levels in SK-BR-3R cells after transfection with MTDH siRNA were detected by Western blotting. Results: Compared to the sensitive SK-BR-3 cells, the sensitivity of SK-BR-3R cells to trastuzumab decreased significantly (P < 0.001), and the expression levels of miR-375 (P < 0.001) and vimentin (P < 0.05) were significantly down-regulated, but the level of EMT marker E-cadherin was significantly up-regulated (P < 0.05). After transfection with miR-375 mimic, the expression levels of miR-375 (P < 0.001) and vimentin (P < 0.01) were up-regulated, while the level of E-cadherin was down-regulated (P < 0.001); the trastuzumab sensitivity of SK-BR-3R cells was increased (P < 0.001). A negative regulatory effect of miR-375 on the transcription of MTDH gene was observed (P < 0.001). After MTDH-siRNA transfection, the expressions of MTDH and vimentin were down-regulated (both P < 0.001), while the expression of E-cadherin was up-regulated (P < 0.001). Conclusion: miR-375 plays a role in drug-resistance of HER2-positive breast cancer cells to trastuzumab through regulating EMT, which is related to the target regulation of MTDH gene expression.

Expressions and prognostic value of metadherin, E-cadherin, and β-catenin in patients with hepatocellular carcinoma / 中华肝胆外科杂志

Objective To study the expressions of metadherin (MTDH),E-cadherin and β-catenin in the tissues of hepatocellular carcinoma (HCC),to investigate the relationship between them and the clinical-pathological features,and to evaluate the prognostic values after surgical resection for hepatocellular carcinoma.Methods The protein expressions of MTDH,E-cadherin and β-catenin were studied by immunohistochemistry in tumor tissues of 107 HCC patients who underwent curative surgical resection.The data were correlated with the clinical-pathological data,tumor free time and recurrence rate.Results Positive expression of MTDH and nuclear β-catenin accumulation were correlated with the Edmondson grade (P＜0.05) and decreased E-cadherin expression was correlated with the preoperative serum level of α-fetoprotein (AFP) (P＜0.05).All these expressions were associated with a shorter tumor-free survival and a higher recurrence rate (P＜0.05).Positive MTDH expression was correlated with decreased E-cadherin expression and nuclear β-catenin accumulation (P＜0.05).On Cox regression analysis,MTDH was an independent risk factor of tumour recurrence (RR=3.431,CI=1.254～ 7.318).Conclusions Positive MTDH expression,decreased E-cadherin expression,and nuclear β-catenin accumulation indicated a higher recurrence rate after curative surgical resection for HCC.MTDH was an independent risk factor of recurrence.

Expression of metadherin in retinoblastoma and its relationship with histopathological high-risk factors / 中华实验眼科杂志

Background Metadherin is a newly discovered oncogene.It is highly expressed in some solid tumors and plays a significant role in invasion and metastasis of neoplasm as an important biological marker of aggressive cancers.However,there is little data available for the relationship between metadherin expression and clinicopathologic features of retinoblastoma(RB).Objective This study was to investigate the expression of the metadherin in RB cell lines and specimens as well as its clinical significance.Methods The expression of metadherin mRNA in different metastatic potential of RB cell lines(Y79,WERI-RB1 and SO-RB50)was detected by real-time PCR,and the protein expression of metadherin (MDTH)in paraffin-embedded RB tissue was detected by immunohistochemistry.The relevance of metadherin expression to clinical histopathological features was statistically analyzed.Results The relative expression value of metadherin mRNA was 6.11±0.17,6.21±0.21 and 3.97±0.17 in Y79,SORB50,WERI-RB1,respectively,with a significant difference among the three types of cell lines(F =142.643,P＜0.05).The relative expression value of metadherin mRNA was significantly higher in Y79 or SO-RB50 than that in W ERI-RB1 (P=0.000).The expression of MTDH protein mostly located in the cellular membrane and cytoplasm.Among the 54 RB sections,35 (64.81％)showed a high intensity in expression of metadherin protein.The total score of metadherin protein expression was higher in the E stage of RB than that of D stage(P=0.035),in the patients with high-risk factor than those without high-risk factor(P=0.002)and the cases with optic nerve invasion compared to non-invasion (P＝0.017).However,no significant differences were found in the expression of metadherin protein between different ages (P =0.579),different genders (P =0.513),bilateral eyes (P =0.305),different disease courses (P=0.860),various types of differentiation (P =0.537),different degree of the ehoroid invasion (P =0.238),and with or without invasion of the anterior ocular segment (P =0.579).Conclusions Metadherin appears to be highly expressed in RB cells.The activation of the MTDH gene is associated with invasion and metastasis of RB.These results imply that the dynamic change of metadherin expression probably is a prognostic indicator of RB.

Metadherin Is a Prognostic Predictor of Hepatocellular Carcinoma after Curative Hepatectomy

BACKGROUND/AIMS: The prognosis after surgical resection of hepatocellular carcinoma (HCC) remains poor because of a high rate of recurrence. Thus, it is crucial to identify patients with a high risk of recurrence after curative hepatectomy and to develop more effective and targeted treatment strategies to improve disease outcomes. In this study, we investigated the roles of metadherin (MTDH) in the prognosis of HCC. METHODS: We investigated MTDH expression using immunohistochemistry in tumor tissue microarrays of 288 primary HCC patients who underwent curative surgical resection. RESULTS: High MTDH expression was observed in 138 of the 288 HCC cases (47.9%). High MTDH expression was associated with a younger age (p<0.001), higher Edmondson grade (p<0.001), microvascular invasion (p<0.001), higher American Joint Committee on Cancer T stage (p=0.001), and higher alpha-fetoprotein level (p=0.003). Multivariate analyses revealed that high MTDH expression (p=0.014), higher Barcelona-Clinic Liver Cancer (BCLC) stage (p<0.001), and Edmondson grade III (p=0.042) were independent predictors of shorter disease-free survival (DFS). Higher BCLC stage (p<0.001) and Edmondson grade III (p=0.047) were also independent predictors of shorter disease-specific survival. CONCLUSIONS: High MTDH expression may be a prognostic predictor of shorter DFS in HCC patients after curative hepatectomy.