作者机构：
[Jiang, H; Su, Q; Zhang, Shuo; Su, Bo; Su, Qi; Zhou, Zhi-Gang; Ling, Hui; Jiang, Hao; Ai, Xiao-Hong; Su, Jian; Yang, Bang-Min; Zeng, Ying; Wu, You-Hua; Xia, Hong; Zeng, Xi; Liu, Fang; Ma, Yan-Hua] Univ South China, Affiliated Hosp 1, Ctr Gastr Canc Res Hunan Prov, Hengyang 421001, Hunan, Peoples R China.;[Zhang, Shuo; Su, Bo; Su, Qi; Zhou, Zhi-Gang; Ling, Hui; Su, Jian; Yang, Bang-Min; Zeng, Ying; Xia, Hong; Zeng, Xi; Liu, Fang; Ma, Yan-Hua] Univ South China, Key Lab Canc Cellular & Mol Pathol Hunan Prov Uni, Canc Res Inst, Hengyang 421001, Hunan, Peoples R China.;[Su, Bo] Univ South China, Inst Pharm & Pharmacol, Key Lab Pharmacoprote Hunan Prov Univ, Hengyang 421001, Hunan, Peoples R China.;[Su, Jian] Univ South China, Affiliated Hosp 2, Dept Pathol, Hengyang 421001, Hunan, Peoples R China.
通讯机构：
[Jiang, H; Su, Q; Su, Qi] U;Univ South China, Affiliated Hosp 1, Ctr Gastr Canc Res Hunan Prov, Hengyang 421001, Hunan, Peoples R China.;Univ South China, Key Lab Canc Cellular & Mol Pathol Hunan Prov Uni, Canc Res Inst, Hengyang 421001, Hunan, Peoples R China.
关键词：
LIMK1;diallyl disulfide;gastric cancer cell epithelial-mesenchymal transition;invasion;proliferation
摘要：
Diallyl disulfide (DADS) has been shown to have multi-targeted antitumor activities. We have previously discovered that it has a repressive effect on LIM kinase-1 (LIMK1) expression in gastric cancer MGC803 cells. This suggests that DADS may inhibit epithelial-mesenchymal transition (EMT) by downregulating LIMK1, resulting in the inhibition of invasion and growth in gastric cancer. In this study, we reveal that LIMK1 expression is correlated with tumor differentiation, invasion depth, clinical stage, lymph node metastasis, and poor prognosis. DADS downregulated the Rac1-Pak1/Rock1-LIMK1 pathway in MGC803 cells, as shown by decreased p-LIMK1 and p-cofilin1 levels, and suppressed cell migration and invasion. Knockdown and overexpression experiments performed in vitro demonstrated that downregulating LIMK1 with DADS resulted in restrained EMT that was coupled with decreased matrix metalloproteinase-9 (MMP-9) and increased tissue inhibitor of metalloproteinase-3 (TIMP-3) expression. In in vitro and in vivo experiments, the DADS-induced suppression of cell proliferation was enhanced and antagonized by the knockdown and overexpression of LIMK1, respectively. Similar results were observed for DADS-induced changes in the expression of vimentin, CD34, Ki-67, and E-cadherin in xenografted tumors. These results indicate that downregulation of LIMK1 by DADS could explain the inhibition of EMT, invasion and proliferation in gastric cancer cells.
摘要：
Transplantation of peripheral blood lymphocytes (PBLs) from healthy humans with latent Epstein-Barr virus (EBV) infection into severe combined immunodeficiency (SCID) mice results in development of EBV-associated human B-cell lymphoma. However, the expression of EBV genes in relation to lymphoma development has not been reported. We investigated latent membrane protein (LMP) and EBV nuclear antigen (EBNA) gene expression in PBLs from EBV-positive blood donors and induced-lymphoma cells from SCID mice to elucidate the functions and effects of the EBV genome in the occurrence and development of lymphoma. PBLs were isolated from 9 healthy blood donors and transplanted into SCID mice. Gene expression levels of LMP-1, LMP-2A, and LMP-2B and EBNA-1, EBNA-2, EBNA-3A, EBNA-3B, EBNA-3C and EBNA-LP were monitored by real-time quantitative-polymerase chain reaction (qRT-PCR) in cells from nine EBV-induced lymphomas and in matched lymphocytes from healthy subjects. LMP-1, EBNA-1 and EBNA-2 protein levels were detected by western blotting. As a result, LMP-1, LMP-2A and LMP-2B mRNA levels were upregulated 256-, 38- and 331-fold, respectively, in the EBV-induced lymphoma cells compared with the controls, while EBNA-1 and EBNA-3A mRNA levels were upregulated 1157- and 1154-fold, respectively. EBNA-2, EBNA-3B, EBNA-3C and EBNA-LP mRNAs were detected in lymphoma cells, but not in lymphocytes from EBV-positive blood donors. LMP-1 and EBNA-2 proteins were not expressed in lymphocytes from EBV-positive blood donors, according to western blotting. Weak EBNA-1 expression was observed in lymphocytes from blood donors with latent EBV infection, while LMP-1, EBNA-1 and EBNA-2 protein levels were significantly upregulated in EBV-induced lymphoma cells, consistent with mRNA expression levels detected by qRT-PCR. In conclusion, LMP-1, LMP-2A, LMP-2B, EBNA-1 and EBNA-3A were upregulated in EBV-induced lymphoma cells, while EBNA-2, EBNA-3B, EBNA-3C and EBNA-LP were absent in lymphocytes from humans with latent EBV infection, but were positively expressed in EBV-induced lymphoma cells.
摘要：
Background: Calcium overload plays an important role in ischemia/reperfusion injury during ischemic brain damage and is mediated by calmodulin (CaM). However, the understanding of the regulatory mechanisms of CaM expression at the gene level is limited. The expression levels of miR-26b change significantly during ACI, and bioinformatic analyses predict that miR-26b would be a potential regulator of calmodulin (CALM1) mRNA. This study aimed to determine the expression of miR-26b and CaM in the plasma of patients with ACI and investigate the impact of miR-26b on CALM1 expression. Methods: CaM and miR-26b expression analyses from the plasma of patients with ACI and normal controls were performed using ELISA and qRT-PCR, respectively. Correlations between CaM, miR-26b, and NIHSS scores were analyzed. Then, miR-26b mimics and inhibitors were transfected into HUVE cell lines via lipofectamine. CALM1 mRNA expression in HUVECs was detected by RT-PCR, and the protein levels were detected by Western blot. Results: Plasma CaM expression in patients with ACI was significantly higher when compared with normal controls, and miR-26b expression was significantly lower. The plasma levels of CaM and miR-26b were correlated with the NIHSS scores in ACI patients. miR-26b modulated CALM1 in vitro. The transfected miR-26b mimic and inhibitor significantly altered the expression of CALM1/CAM at the mRNA and protein levels in cultured HUVECs. Conclusions: CaM might be a potential novel blood marker in patients with ACI. miR-26b targeted CALM1 and affected the expression of CaM at the post-transcriptional level, which likely contributed to the progression of ACI brain injury.
摘要：
Ur anium release into the environment is a threat to human health, and the mechanisms of cytotoxicity caused by uranium are not well-understood. To improve our understanding in this respect, we herein evaluated the effects of uranium exposure on normal rat hepatic BRL cells. As revealed by scanning electron microscopy and transmission electron microscope analysis, uranyl nitrate was found to be transformed into uranyl phosphate particles in the medium and taken up by BRL cells in an endocytotic uptake manner, which presumably initiates apoptosis of the cell, although soluble uranyl ion may also be toxic. The apoptosis of BRL cells upon uranium exposure was also confirmed by both the acridine orange and ethidium bromide double staining assay and the Annexin V/propidium iodide double staining assay. Further studies revealed that uranium induced the loss of mitochondrial membrane potential in a dose-dependent manner. Moreover, the uranium-induced apoptosis was found to be associated with the activation of caspase-3, caspase-8 and caspase-9, indicating both a mitochondria-dependent signaling pathway and a death receptor pathway by a crosstalk. This study provides new chemical and biological insights into the mechanism of uranium toxicity toward hepatic cells, which will help seek approaches for biological remediation of uranium.
