MicroRNA-877-5p is involved in the trovafloxacin-induced liver injury.
Mitsugi Ryo,Itoh Tomoo,Fujiwara Ryoichi
Trovafloxacin develops severe hepatotoxicity; however, the underlying mechanism of the trovafloxacin-induced liver injury has not been cleared. It has been shown that microRNAs (miRNAs) can be involved in the development of drug-induced liver injuries. We performed a miRNA microarray analysis to identify hepatic miRNAs that were induced or reduced by trovafloxacin in mice. It was demonstrated that miR-877-5p was the most increased miRNA in the mouse liver 24h after the trovafloxacin administration. To investigate the role of miR-877-5p in the liver, we established miR-877-5p-overexpressed HepG2 cells. Microarray analysis detected altered expressions in 2077 (>2-fold) and 1547 (<0.5-fold) genes in the miR-877-5p overexpressing cells compared to the mock cells. Especially, SLCO4C1, PEPCK, MT1M, HIST1H2BM, LGI1, and PLA2G2A were markedly increased or decreased in the miR-877-5p overexpressing cells. We conducted a correlation analysis between the expression levels of miR-877-5p and the six genes in eight miR-877-5p stably-expressed clones. It was shown that the PEPCK expression levels were correlated with miR-877-5p expression levels. PEPCK is associated with development of apoptotic cell death; therefore, the increased miR- 877-5p-induced PEPCK can be a trigger that is involved in the development of trovafloxacin-induced liver injury.
Expression, regulation and function of human metallothioneins in endothelial cells.
Schulkens Iris A,Castricum Kitty C M,Weijers Ester M,Koolwijk Pieter,Griffioen Arjan W,Thijssen Victor L
Journal of vascular research
Metallothioneins (MTs) are small cysteine-rich proteins which are involved in e.g. metal homeostasis, metal detoxification and protection against oxidative stress. In addition, several MTs have been shown to regulate expression of proangiogenic growth factors like vascular endothelial growth factor. Detailed information about the expression and regulation of specific MT isoforms in endothelial cells (EC) is limited. We therefore performed extensive mRNA expression profiling of all known human MTs in EC. We found that the basal endothelial expression is restricted to MT1E, MT1X, MT2A, and MT3. Physiological activation of EC by exposure to serum increased the expression of MT1E and MT2A and induced the expression of MT1M. Furthermore, exposure to zinc or copper induced the expression of most MT1 isoforms, while hypoxia specifically increased the expression of MT1E, MT1M, MT1X, and MT3. Finally, knockdown of the dominant MT isoform in EC, i.e. MT2A, resulted in decreased proliferation and sprouting as well as in increased migration of human umbilical vein EC. Together, these findings provide a link between MTs and angiogenesis.
MiR-24-3p enhances cell growth in hepatocellular carcinoma by targeting metallothionein 1M.
Dong Xiaogang,Ding Wei,Ye Jianwei,Yan Dong,Xue Feng,Xu Lin,Yin Jiwei,Guo Wenjia
Cell biochemistry and function
Dysregulation of microRNAs has been demonstrated to contribute to malignant progression of cancers, including hepatocellular carcinoma (HCC). MiR-24-3p was previously reported to be significantly upregulated in HCC. However, the potential role and mechanism of action of miR-24-3p in the initiation and progression of HCC remain largely unknown. Quantitative reverse transcription polymerase chain reaction demonstrated that miR-24-3p was significantly upregulated in HCC tumor tissues compared with nontumor tissues. The cell viability, colony formation assay, and tumorigenicity assays in nude mice showed that miR-24-3p could enhance HCC cell growth in vitro and in vivo. Metallothionein 1M was verified as an miR-24-3p target gene by using dual-luciferase reporter assays, quantitative reverse transcription polymerase chain reaction, and Western blotting, which was involved in miR-24-3p regulated HCC cell growth. These results indicated that miR-24-3p plays an important role in the initiation and progression of HCC by targeting metallothionein 1M, and the miR-24-3p/metallothionein 1M pathway may contribute to the development of novel therapeutic strategies for HCC in the future.
The roles of metallothioneins in carcinogenesis.
Si Manfei,Lang Jinghe
Journal of hematology & oncology
Metallothioneins (MTs) are small cysteine-rich proteins that play important roles in metal homeostasis and protection against heavy metal toxicity, DNA damage, and oxidative stress. In humans, MTs have four main isoforms (MT1, MT2, MT3, and MT4) that are encoded by genes located on chromosome 16q13. MT1 comprises eight known functional (sub)isoforms (MT1A, MT1B, MT1E, MT1F, MT1G, MT1H, MT1M, and MT1X). Emerging evidence shows that MTs play a pivotal role in tumor formation, progression, and drug resistance. However, the expression of MTs is not universal in all human tumors and may depend on the type and differentiation status of tumors, as well as other environmental stimuli or gene mutations. More importantly, the differential expression of particular MT isoforms can be utilized for tumor diagnosis and therapy. This review summarizes the recent knowledge on the functions and mechanisms of MTs in carcinogenesis and describes the differential expression and regulation of MT isoforms in various malignant tumors. The roles of MTs in tumor growth, differentiation, angiogenesis, metastasis, microenvironment remodeling, immune escape, and drug resistance are also discussed. Finally, this review highlights the potential of MTs as biomarkers for cancer diagnosis and prognosis and introduces some current applications of targeting MT isoforms in cancer therapy. The knowledge on the MTs may provide new insights for treating cancer and bring hope for the elimination of cancer.
lncRNA STEAP3-AS1 Modulates Cell Cycle Progression via Affecting CDKN1C Expression through STEAP3 in Colon Cancer.
Na Heya,Li Xiaomeng,Zhang Xinsheng,Xu Yue,Sun Yuzhu,Cui Jingyi,Chen Zihao,Shi Xiaomeng,Ren Shuangyi,Zuo Yunfei
Molecular therapy. Nucleic acids
Previous studies have reported that long noncoding RNAs (lncRNAs) have acted as new players during tumorigenesis. Metallothionein also plays an important role in tumor progression. It is mainly considered to be involved in the process of cell proliferation, oxidative stress, and multidrug resistance. However, the potential involvement of metallothionein-related lncRNAs in colon cancer remains poorly understood. In our study, we found that MT1M affected the expression of lncRNA STEAP3-AS1. STEAP3-AS1 is located in physical contiguity with STEAP3 and notably increased in colon cancer tissues and cell lines. STEAP3-AS1 expression was negatively associated with the expression of STEAP3. High levels of STEPA3-AS1 were associated with poor overall survival in colon cancer patients. In in vitro assays, STEAP3-AS1 knockdown could inhibit colon cancer cell proliferation and migration and arrest colon cancer cells at the G-G phase. In tumorigenicity assays, STEAP3-AS1 knockdown could strongly inhibit tumor growth. Mechanistic investigations demonstrated that STEAP3-AS1 downregulation could increase the expression of cyclin-dependent kinase inhibitor 1C (CDKN1C) by STEAP3 upregulation. Overall, we identify the underlying role of MT1M-related lncRNA STEAP3-AS1 in colon cancer progression, which provides a novel strategy for colon cancer therapy.
[Effect of MT1M gene on the cell cycle and signaling pathway of Hep-G2].
Sun Luhong,Zhang Xiran,Kong Yahui,Yu Long
Zhonghua nan ke xue = National journal of andrology
OBJECTIVE:To investigate the possible roles of MT1M gene on the cell cycle and signaling pathway of Hep-G2. METHODS:Hep-G2 human hepatoma cells made by transfection with expressible MT1M gene, and the cell cycle was detected by flow cytometry, and the signaling pathway was measured by dual luciferase assay in Hep-G2 cells. RESULTS:MT1M gene was able to induce changes of the cell cycle and the activation of NF-kappaB pathway in Hep-G2 cells. CONCLUSION:MT1M gene may affect the cell cycle in Hep-G2 and activate the NF-kappaB-dependent transcription.
MT1M and MT1G promoter methylation as biomarkers for hepatocellular carcinoma.
Ji Xiang-Fen,Fan Yu-Chen,Gao Shuai,Yang Yang,Zhang Jian-Jun,Wang Kai
World journal of gastroenterology
AIM:To investigate the potential of promoter methylation of two tumor suppressor genes (TSGs) as biomarkers for hepatocellular carcinoma (HCC). METHODS:A total of 189 subjects were included in this retrospective cohort, which contained 121 HCC patients without any history of curative treatment, 37 patients with chronic hepatitis B (CHB), and 31 normal controls (NCs). DNA samples were extracted from 400 μL of serum of each subject and then modified using bisulfite treatment. Methylation of the promoters of the TSGs (metallothionein 1M, MT1M; and metallothionein 1G, MT1G) was determined using methylation-specific polymerase chain reaction. The diagnostic value of combined MT1M and MT1G promoter methylation was evaluated using the area under the receiver operating characteristic curves. RESULTS:Our results indicated that the methylation status of serum MT1M (48.8%, 59/121) and MT1G (70.2%, 85/121) promoters in the HCC group was significantly higher than that in the CHB group (MT1M 5.4%, 2/37, P < 0.001; MT1G 16.2%, 6/37, P < 0.001) and NC group (MT1M 6.5%, 2/31, P < 0.001; MT1G 12.9%, 4/27, P < 0.001). Aberrant serum MT1M promoter methylation gave higher specificity to discriminate HCC from CHB (94.6%) and NCs (93.5%), whereas combined methylation of serum MT1M and MT1G promoters showed higher diagnostic sensitivity (90.9%), suggesting that they are potential markers for noninvasive detection of HCC. Furthermore, MT1M promoter methylation was positively correlated with tumor size (rs = 0.321, P < 0.001), and HCC patients with both MT1M and MT1G promoter methylation tended to show a higher incidence of vascular invasion or metastasis (P = 0.018). CONCLUSION:MT1M and MT1G promoter methylation may be used as serum biomarkers for noninvasive detection of HCC.
Metallothionein 1M suppresses tumorigenesis in hepatocellular carcinoma.
Fu Cheng-Lin,Pan Bing,Pan Ju-Hua,Gan Mei-Fu
Members of the metallothionein (MT) family are involved in metal detoxifcation and in the protection of cells against certain electrophilic carcinogens. In present study, it was found that MT1M was downregulated in more than 77.1% (91/118) of hepatocellular carcinoma (HCC) tissues compared with adjacent non-tumor tissues. Furthermore, overexpression of MT1M inhibited cell viability, colony formation, cell migration and invasion in HCC cell lines and tumor cell growth in xenograft nude mice, and activated cell apoptosis in HCC cell lines. In addition, immunohistochemistry analysis showed MT1M was negative or weak staining in tumor tissues but moderate or strong staining in adjacent non-tumor tissues. The sensitivity and specificity of MT1M for HCC diagnosis were 76.27% and 89.83%, respectively. In conclusion, MT1M was identified as a potential tumor marker for HCC and may serve as a useful therapeutic agent for HCC gene therapy.
The effect and mechanism of metallothionein MT1M on hepatocellular carcinoma cell.
Zhang S,Huang Z,Zhou S,Wang B,Ding Y,Chu J-Z,Wang X-L
European review for medical and pharmacological sciences
OBJECTIVE:Liver cancer is one of the most common digestive system malignant solid tumors. Its incidence and mortality rates keep high, causing serious mental and economic burden. So far, the exact mechanism of liver cancer onset has not been fully elucidated. Metallothionein (MT) widely exists in various types of organisms with highly conserved structure. It contains the short peptide of cysteine and sulfur protein with high affinity to heavy metals, including cadmium, zinc, and copper. MT1M is an important member of MT family that has been verified to participate in regulating hepatocellular carcinoma, thyroid cancer, cervical cancer, and other tumors. However, MT1M expression and mechanism in hepatoma cells have not been fully elucidated. MATERIALS AND METHODS:Hepatoma cell line HepG2 was divided into control and MT1M group. MT1M plasmid was constructed and transfected to MT1M group. Real-time PCR was used to test MT1M expression. MTT assay was applied to detect HepG2 proliferation. Flow cytometry was performed to determine HepG2 apoptosis. Caspase-3 activity was measured. Western blot was used to detect Bcl-2 and Bax protein levels. RESULTS:MT1M expression significantly increased after MT1M plasmid transfection compared with control (p < 0.05). MT1M group showed inhibited HepG2 proliferation, declined HepG2 apoptosis, enhanced Caspase-3 activity, reduced Bcl-2 protein level, and upregulated Bax protein compared with control (p < 0.05). CONCLUSIONS:MT1M can suppress HepG2 proliferation and induce HepG2 apoptosis through downregulating Bcl-2, upregulating Bax, and enhancing Caspase-3 activity.
Low metallothionein 1M (MT1M) is associated with thyroid cancer cell lines progression.
Chen Yizuo,Quan Ruida,Bhandari Adheesh,Chen Zheng,Guan Yaoyao,Xiang Jingjing,You Jie,Teng Lisong
American journal of translational research
Papillary thyroid cancer (PTC) is the most common malignancy of the thyroid carcinoma, despite ongoing advances, novel biomarkers are required for prognosis and diagnosis of PTC. Our previous research found that metallothionein 1M (MT1M) was a novel potential PTC associated gene in thyroid cancer. In the present study, the expression status and prognostic value of MT1M expression were investigated in thyroid cancer. Tissue samples from 60 patients with PTC were subjected to the quantitative real-time polymerase chain reaction and the relative expression of MT1M in the patient tissue was evaluated. The Cancer Genome Atlas (TCGA) RNA-seq database was downloaded to further explore the role of MT1M in PTC and its relationship with lymph node metastasis (LNM). Logistic analysis showed that reduced expression of MT1M, histological type, and clinical stage are independent high-risk factors for LNM in PTC. The biological function of MT1M was also researched by using the PTC cell lines TPC-1, KTC1 and BCPAP. In vitro experiments revealed that MT1M upregulation significantly inhibits the colony formation, proliferation, migration, and invasion of PTC cell lines. We also found that MT1M could modulate the expression of N-cadherin and vimentin. These results implied that MT1M involved in the progress of thyroid cancer and might act as a tumor suppressor gene. In this study, we identified, for the first time, MT1M was involved in thyroid carcinoma cell lines This study specified a potential new marker and a target for gene therapy in thyroid cancer treatment.
MiR-545-3p/MT1M axis regulates cell proliferation, invasion and migration in hepatocellular carcinoma.
Changjun Liu,Feizhou Huang,Dezhen Peng,Zhao Lei,Xianhai Mao
Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie
Studies have shown that metallothionein 1 M (MT1M) is a tumor suppressor gene which is frequently down-regulated in human hepatocellular carcinoma (HCC). The methylation of MT1M promoter region is one of the important transcriptional regulation mechanisms that contribute to the loss of its expression. In our study, we found that there are still half of the 55 HCC tumor tissues in our cohort do not share the promoter methylation of MT1M. So, we speculated there maybe another mechanism participating in the downregulation of MT1M in HCC. Then, we provided evidences that miR-545-3p, which served as a tumor promoter, post-transcriptionally regulate MT1M in HCC through binding to its untranslated region (3'UTR). Taking together, we investigated the role of miR-545-3p in the process of HCC through regulating MT1M.
Metallothionein 1M (MT1M) inhibits lung adenocarcinoma cell viability, migration, and expression of cell mobility-related proteins through MDM2/p53/MT1M signaling.
Translational cancer research
BACKGROUND:Metallothionein 1M (MT1M) functions to regulate cell proliferation and cancer metastasis. This study assessed the effects of MT1M overexpression and mouse double minute 2 homolog (MDM2) knockdown on the regulation of non-small cell lung cancer A549 cell viability, migration, and protein expression and explored the underlying molecular events. METHODS:A549 cells were stably infected with lentivirus carrying MT1M cDNA or transiently transfected MDM2 siRNA and/or treated with the p53 inhibitor for the assessment of changes in cell viability, wound healing, Transwell migration, and qRT-PCR and Western blot assays. Luciferase reporter assay was performed to investigate p53 binding to the MT1M promoter. RESULTS:The data showed that MT1M overexpression inhibited A549 cell viability and migration capacity , whereas the p53 inhibitor reversed the inhibition of A549 cell viability and migration caused by MT1M overexpression as well as the expression of MMP2, MMP9, and MMP14. Furthermore, knockdown of MDM2, an upstream inhibitor of p53 activity, was able to reduce A549 cell viability, migration, and protein expression. Thus, MDM2 knockdown had synergistic effects with MT1M overexpression on the suppression of A549 cell viability, migration, and protein expression. CONCLUSIONS:In conclusion, MDM2 can bind to and phosphorylate p53 protein to inactivate the protein, thereby reducing MT1M expression and leading to tumor cell proliferation and migration.
MTIM affects retinal ganglion cells through PI3K/AKT signaling pathway.
Li R-X,Wang Q,Shen Z-W,Jie L-M
European review for medical and pharmacological sciences
OBJECTIVE:Retinal ganglion cells (RGCs) are the main cells that form vision in the retina. MT1M is involved in the occurrence and progression of various diseases. However, the role of MTIM in RGCs cells remains unclear. MATERIALS AND METHODS:RGCs were cultured in vitro and randomly divided into control group, MT1M group (transfected with MT1M-pcDNA3.1 plasmid), and MT1M siRNA group (transfected with MT1M siRNA) followed by measuring MT1M and NT-3 expression by real time PCR and Western blot, cell proliferation by MTT assay, secretion of IL-2 and IL-6 by enzyme-linked immunosorbent assay (ELISA), SOD activity and ROS content. In addition, expression of PI3K/AKT signaling pathway protein was detected by Western blot. RESULTS:MT1M expression in MT1M group was significantly increased, which promoted cell proliferation, increased NT-3 expression, and decreased Caspase 3 activity and IL-2 and IL-6 secretion. Meanwhile, SOD activity was increased, ROS content was decreased and PI3K/AKT protein phosphorylation was elevated. The differences were statistically significant compared with control group (p < 0.05). MT1M siRNA decreased MT1M expression, inhibited cell proliferation, decreased NT-3, and increased Caspase 3 activity and IL-2 and IL-6 secretion. In addition, MT1M siRNA decreased SOD activity, increased ROS content and reduced PI3K/AKT protein phosphorylation. Compared with control group, the differences were statistically significant (p < 0.05). CONCLUSIONS:Up-regulation of MT1M can inhibit RGC cell apoptosis and inflammation, and promote RGC cell proliferation through the PI3K/AKT signaling pathway.