Effect of Copper-Containing Stainless Steel on Apoptosis of Coronary Artery Smooth Muscle Cells.

BACKGROUND: We aimed to investigate the effect of copper stainless steel on apoptosis of vascular smooth muscle cells in coronary artery. METHODS: The study was carried out in 2019 at Hubei University of Medicine, Xiangyang, China. The rat coronary artery smooth muscle cell was used for cell resuscitation and culture. MTT method was used to visualize cell growth curve and to detect the cell survival and growth. The incubated cells were randomly divided into copper-containing stainless-steel group, ordinary stainless-steel group, and control group. The cells were made into single cell suspension, which were intervened by experimental group and incubated in incubator with CO2 for 48 hours. TUNEL method was used to detect the apoptosis. The number of apoptotic cells in five high power fields (×200) was counted. The expression of Fas protein in three groups of cells was detected by Western blot. RESULTS: The growth curves of rat coronary artery smooth muscle cells showed that the OD value of the cells reached the plateau 7 days after inoculation, indicating that the cells grew well. TUNEL staining showed the apoptosis in all three groups. The apoptotic index in copper-containing group was significantly higher than that in common stainless-steel group (P <0.01). The results of the Fas protein expression level through Western blot showed that the level in the copper-containing group was significantly higher than that in the common stainless-steel group (P<0.01). CONCLUSION: Copper-containing stainless steel can promote apoptosis of coronary artery smooth muscle cells. The material could prevent stent restenosis.

Long non-coding RNA ADAMTS9-AS2 inhibits liver cancer cell proliferation, migration and invasion.

Long non-coding RNA (lncRNA) ADAM metallopeptidase with thrombospondin type 1 motif 9 antisense RNA 2 (ADAMTS9-AS2) is involved in various types of cancer, such as ovarian cancer, lung cancer and clear cell renal cell carcinoma. However, the roles of ADAMTS9-AS2 in liver cancer are not completely understood. The present study aimed to determine the functional role of ADAMTS9-AS2 in human liver cancer and investigate the potential underlying molecular mechanisms. The expression levels of ADAMTS9-AS2 and ADAMTS9 were determined following ADAMTS9-AS2 overexpression and knockdown. The results indicated that ADAMTS9-AS2 overexpression and knockdown increased and decreased ADAMTS9 mRNA and protein expression levels, respectively, indicating that alterations in ADAMTS9 expression corresponded with ADAMTS9-AS2 expression. Subsequently, the effects of ADAMTS9-AS2 on liver cancer cell proliferation, migration and invasion were analyzed by performing Cell Counting Kit-8, wound healing and Transwell assays, respectively. The results demonstrated that ADAMTS9-AS2 inhibited liver cancer cell proliferation, migration and invasion. Finally, the effect of ADAMTS9 on PI3K/AKT/mTOR signaling pathway-associated proteins [AKT, phosphorylated-AKT, phosphatidylinositol-4, 5-bisphosphate 3-kinase catalytic subunit ß (PIK3CB), mTOR and phosphorylated-mTOR], several key autophagy-related proteins [light chain 3-I/II (LC3-I/II), beclin 1 (BECN1) and sequestosome 1 (SQSTM1)] and apoptosis-related proteins (Bax and Bcl-2) was detected via western blotting. The results suggested that ADAMTS9-AS2 downregulated the phosphorylation of AKT and mTOR, the protein expression level of PIK3CB, as well as the expression levels of autophagy protein SQSTM1 and antiapoptotic protein Bcl-2. By contrast, ADAMTS9-AS2 upregulated the expression levels of autophagy proteins LC3-II and BECN1, and the proapoptotic protein Bax. Collectively, ADAMTS9-AS2 inhibited liver cancer cell proliferation, migration and invasion via inhibiting the PI3K/AKT/mTOR signaling pathway. The present study provided a novel insight into the role of ADAMTS9-AS2 in liver cancer.

miR-144 suppresses the proliferation and metastasis of hepatocellular carcinoma by targeting E2F3.

MicroRNAs (miRNAs) play essential roles in the progression of hepatocellular carcinoma (HCC). miR-144 acts as a tumor suppressor in some malignancies, while its role in HCC is unclear. Here, we found that miR-144 was significantly decreased in HCC tissues and cell lines. Forced overexpression of miR-144 remarkably reduced cell proliferation, increased apoptosis, and suppressed migration and invasion of HCC cells. E2F transcription factor 3 (E2F3) was identified as a target of miR-144 in HCC cells. Moreover, E2F3 overexpression partially attenuated the tumor suppressive effects of miR-144, and the expression of E2F3 was negatively correlated with miR-144 level in HCC tissues. Our data suggest that miR-144 might suppress the growth and motility of HCC cells partially by targeting E2F3.

[Expression of ADAMTS-2 and TGF-ß1 in cirrhotic liver].

OBJECTIVE: To explore the expression and distribution of a disintegrin and metalloprotease with thrombospondin motif (ADAMTS)-2 and transforming growth factor (TGF) -ß1 in patients with or without cirrhosis, and to determine their relation. METHODS: The liver tissues from 16 patients with cirrhotic portal hypertensive and 8 patients with liver injury were collected in Wuhan General Hospital from March to June, 2010. Immunohistochemistry and Western blot were applied to detect the protein expression of ADAMTS-2 and TGF-ß1. RESULTS: Immunohistochemistry showed that the expression of ADAMTS-2 and TGF-ß1 was significantly higher in the cirrhotic tissues than that in normal tissues (P<0.05). Western blot also showed the expression of ADAMTS-2 and TGF-ß1 in the cirrhosis tissues was significantly higher than that in normal tissues (P<0.05). There was a positive correlation between ADAMTS-2 and TGF-ß1 (r=0.862, P<0.01). CONCLUSION: ADAMTS-2 and TGF-ß1 may have a synergistic reaction in promoting liver cirrhosis.

[Significances of gene differential expression patterns in hepatocirrhosis and non-hepatocirrhosis tissues within different ischemic time].

OBJECTIVE: To investigate the gene differential expression patterns in hepatocirrhosis and non-hepatocirrhosis tissues within different ischemic time. METHODS: The liver tissues were divided into two groups: Group A (non-hepatocirrhosis), Group B (hepatocirrhosis), each of which consisted of 3 groups with different ischemic time: 15, 30 and 45 minutes. The gene differential expression patterns in the two groups within different ischemic time were detected and compared with those in normal liver tissues by using 4000 points gene microarray. RESULTS: In non-hepatocirrhosis tissues, the homeostatic maintenance genes expressed highly during hepatic ischemia for 15 minutes, and no apoptotic gene was expressed; but in hepatocirrhosis tissues, many apoptotic genes expressed highly. As for 30 minutes, in both two groups liver tissue genes expressed to the peak, and the genes related to cell death, oxidative stress and nuclear factors expressed highly. The difference lies in the facts that in Group B pro-apoptosis genes expressed more than those in Group A, and the Ratio values were higher than those in Group A. Many genes of heat shock protein family and antioxidant proteins expressed highly simultaneously in Group A, but comparatively low in Group B. As for 45 minutes, genes of heat shock proteins and antioxidant proteins expressed lowly in Group B. CONCLUSIONS: It suggests that the safe time limit of hepatic ischemia for cell survive is 30 minutes or so. Non-hepatocirrhosis tissues could endure 30 minutes of ischemia and even longer, but it should be restricted within 30 minutes in hepatocirrhosis tissues.

Multiple gene differential expression patterns in human ischemic liver: safe limit of warm ischemic time.

AIM: To investigate the multiple gene differential expression patterns in human ischemic liver and to produce the evidence about the hepatic ischemic safety time. METHODS: The responses of cells to hepatic ischemia and hypoxia at hepatic ischemia were analyzed by cDNA microarrary representing 4 000 different human genes containing 200 apoptotic correlative genes. RESULTS: There were lower or normal expression levels of apoptotic correlative genes during the periods of hepatic ischemia for 0-15 min, the maintenance homostatic genes were expressed significantly higher at the same time. But at the hepatic ischemia for 30 min, the expression levels of maintenance homeostatic genes were down-regulated, the expressions of many apoptotic correlative genes and nuclear transcription factors were activated and up-regulated. CONCLUSION: HIF-1, APAF-1, PCDC10, FBX5, DFF40, DFFA XIAP, survivin may be regarded as the signal genes to judge the degree of hepatic ischemic-hypoxic injure, and the apoptotic liver cell injury due to ischemia in different time limits. The safe limit of human hepatic warm ischemic time appears to be generally less then 30 min.