The function of Trim72 gene in hepatocellular carcinoma metastasis / 国际生物医学工程杂志

Objective To investigate the role of non-small cell lung cancer metastasis-related Trim72 gene in hepatocellular carcinoma (HCC) using clustered regularly interspaced short palindromic repeats (CRISPR) Cas9 system.Methods Hepa1-6 (Cas9) HCC cells were established with stable expression of Cas9 protein,and then specific gene knockout was performed using sgRNA targeting Trim72 gene.After obtaining the Hepa1-6 (Trim72-KO) cells,the metastasis and invasion abilities of the cells were evaluated by in vitro Transwell assay and in vivo subcutaneous lung metastasis examination.Results Hepa1-6 (Trim72-KO) cell line was successfully established by the CRISPR-Cas9 system.Transwell assay indicated that the mobility of Hepa1-6 (Trim72-KO) cells was increased compared to the control cells.Transwell assay indicated that the metastasis and invasion of Hepa1-6 (Cas9) HCC cells were enhanced after the knockout of Trim72 gene.The pathological examination of lung metastasis of subcutaneous tumor in vivo showed that the subcutaneously metastatic ability of Hepa1-6 (Trim72-KO) cells (the experimental group) was significantly stronger than Hepa1-6 (Cas9) cells (the control troup) that were not transferred to the corresponding sgRNA.Conclusions The trim72 gene knocked-out HCC cells were obtained by CRISPR-Cas9 system,which showed stronger metastasis and invasion abilities than the control cells.It is suggested that Trim72 gene may play an important role in the invasion and metastasis of HCC,and Trim 72 gene is expected to be a potential target for gene therapy of liver cancer.

Expression levels of sarcolemmal membrane repair proteins following prolonged exercise training in mice.

Membrane repair is a conserved cellular process, where intracellular vesicles translocate to sites of plasma membrane injury to actively reseal membrane disruptions. Such membrane disruptions commonly occur in the course of normal physiology, particularly in skeletal muscles due to repeated contraction producing small tears in the sarcolemmal membrane. Here, we investigated whether prolonged exercise could produce adaptive changes in expression levels of proteins associated with the membrane repair process, including mitsugumin 53/tripartite motif-containing protein 72 (MG53/TRIM72), dysferlin and caveolin-3 (cav3). Mice were exercised using a treadmill running protocol and protein levels were measured by immunoblotting. The specificity of the antibodies used was established by immunoblot testing of various tissue lysates from both mice and rats. We found that MG53/TRIM72 immunostaining on isolated mouse skeletal muscle fibers showed protein localization at sites of membrane disruption created by the isolation of these muscle fibers. However, no significant changes in the expression levels of the tested membrane repair proteins were observed following prolonged treadmill running for eight weeks (30 to 80 min/day). These findings suggest that any compensation occurring in the membrane repair process in skeletal muscle following prolonged exercise does not affect the expression levels of these three key membrane repair proteins.