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Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Difficult‐to‐express (DTE) recombinant proteins such as multi‐specific proteins, DTE monoclonal antibodies, and lysosomal enzymes have seen difficulties in manufacturability using Chinese hamster ovary (CHO) cells or other mammalian cells as production platforms. CHO cells are preferably used for recombinant protein production for their ability to secrete human‐like recombinant proteins with posttranslational modification, resistance to viral infection, and familiarity with drug regulators. However, despite huge progress made in engineering CHO cells for high volumetric productivity, DTE proteins like recombinant lysosomal sulfatase represent one of the poorly understood proteins. Furthermore, there is growing interest in the use of microRNA (miRNA) to engineer CHO cells expressing DTE proteins to improve cell performance of relevant bioprocess phenotypes. To our knowledge, no research has been done to improve CHO cell production of DTE recombinant lysosomal sulfatase using miRNA. We identified miR‐23a and miR‐377 as miRNAs predicted to target SUMF1, an activator of sulfatases, using in silico prediction tools. Transient inhibition of CHO endogenous miR‐23a/miR‐377 significantly enhanced recombinant sulfatase enzyme‐specific activity by ~15–21% compared to scramble without affecting cell growth. Though inhibition of miR‐23a/miR‐377 had no significant effect on the mRNA and protein levels of SUMF1, overexpression of miR‐23a/377 caused ~30% and ~27–29% significant reduction in endogenous SUMF1 protein and mRNA expression levels, respectively. In summary, our data demonstrate the importance of using miRNA to optimize the CHO cell line secreting DTE recombinant lysosomal sulfatase. [ABSTRACT FROM AUTHOR]

Objective: To study the effect of MiR-377-mediated Syk on atherosclerosis in ApoE/mice. Methods: The experimental subjects were divided into control group (ApoE_/mice), MiR- 377 group (ApoE_/mice injected with MiR-377), Syk group (ApoE_/mice injected with Syk) and Syk+ MiR-377 group (ApoE_/mice injected with MiR-377and Syk). MiR-377 singlestranded antisense nucleotide drugs and Syk gene sequences were prepared and transfected into ApoE_/mice. The expression of Syk gene and protein in ApoE_/mice was detected by qPCR and Western blot, and the levels of inflammatory factors, lipid and oxidative dismutase were detected. The effects of MiR-377 on atherosclerotic aortic plaque and surface area in ApoE_/mice were studied by HE staining and oil red O staining. Results: The relative expression of Syk gene in MiR-377 group was significantly lower than that in control group, and the expression of Syk protein in MiR-377 group was significantly lower than that in control group. TNF-alpha levels in Syk group were significantly lower than those in control group and MiR-377 group. The 1L-6 level in the MiR-377 group was significantly higher than that in the control group, while the 1L-6 level in the Syk group and the MiR377+Syk group was significantly lower than that in the control group. The level of 1L-1ß in the MiR377 group was significantly lower than that in the control group. The TC level of MiR-377 group was significantly lower than that of control group, and the 1L-1ß level of Syk group was significantly higher than that of control group and MiR-377 group. The level of LDL in the MiR-377 group was significantly lower than that in the control group, the level of HDL in the MiR-377 group was significantly lower than that in the control group, and the level of HDL in the Syk group was significantly higher than that in the control group and the MiR-377 group. The level of SOD in the MiR-377 group was significantly higher than that in the control group. There were obvious aortic plaques in the control group and Syk group. There were no obvious aortic plaques in the MiR-377+Syk group and the MiR-377 treated group. The area of atherosclerotic plaque in ApoE_/mice treated with MiR-377 decreased significantly. The plaque area of ApoE-/- mice treated with Syk increased. The ratio of sclerotic plaque area to total area was 25.63% in control group, 7.81% in mi-377 group, 35.19% in Syk group and 9.73% in Syk + MiR-377 group. Conclusion: Overexpression of Syk gene can increase lipid accumulation and oxidative stress in atherosclerotic cells. Mir-377 can regulate the development of atherosclerosis by inhibiting the expression of Syk gene. [ABSTRACT FROM AUTHOR]

Accumulating evidence suggests that cardiomyocyte autophagy is relevant to the onset of cardiac hypertrophy (CH). Several miRNAs are involved in the occurrence of heart failure, and relevant therapeutic treatments are currently being developed. MicroRNA-377 (miR-377) is known to correlate with the progression of various cancers, however, its function in CH has not been determined. Therefore, this study aimed to evaluate miR-377 expression in H2C9 hypertrophic cardiomyocytes in vitro and in a murine model of CH. Gene expression changes were verified via qRT-PCR. Western blotting was used for evaluation of alterations in the expression of signaling pathway-related proteins. Our results indicated that miR-377 expression was markedly upregulated in mice with hypertrophic cardiomyopathy. Autophagy markers were downregulated in these mice and in hypertrophic cardiomyocytes following miR-377 transfection. In addition, we demonstrated that miR-377 acts by targeting peroxisome proliferator-activated receptor γ (PPARγ). PPARγ overexpression promoted autophagy and suppressed cyclosporine A-induced CH. In contrast, PPARγ knockdown further suppressed CH and autophagy. In conclusion, our findings indicated that miR-377 accelerates CH by inhibiting autophagy via targeting PPARγ. This newly identified miR-377/PPARγ axis improves our understanding of the molecular mechanisms underlying CH, and provides a potential new therapeutic target for its treatment. [ABSTRACT FROM AUTHOR]

Aberrantly expressed microRNAs (miRNAs/miRs) and their role in cancer development have recently gained more attention. However, the potential role of miRNAs in hepatocellular carcinoma (HCC) remains largely unknown. In this study, we demonstrated that miR-377 was markedly downregulated in HCC cell lines and primary human HCC tissues. The decreased expression of miR-377 contributes to the upregulation of Bcl-xL expression by targeting its 3-untranslated region (3-UTR). Functionally, knockdown of miR-377 noticeably increased HCC cell growth and colony formation and inhibited apoptosis. In contrast, overexpression of miR-377 suppressed cell proliferation and increased apoptosis. This study provides new insights for the use of miR-377 as a potential molecular target in HCC therapy. [ABSTRACT FROM AUTHOR]

High dietary fructose is an important causative factor in the development of metabolic syndrome-associated glomerular podocyte oxidative stress and injury. Here, we identified microRNA-377 (miR-377) as a biomarker of oxidative stress in renal cortex of fructose-fed rats, which correlated with podocyte injury and albuminuria in metabolic syndrome. Fructose feeding increased miR-377 expression, decreased superoxide dismutase (SOD) expression and activity, and caused O 2 − and H 2 O 2 overproduction in kidney cortex or glomeruli of rats. This reactive oxygen species induction increased p38 MAPK phosphorylation and thioredoxin-interacting protein (TXNIP) expression and activated the NOD-like receptor pyrin domain-containing 3 (NLRP3) inflammasome to produce interleukin-1β in kidney glomeruli of fructose-fed rats. These pathological processes were further evaluated in cultured differentiated podocytes exposed to 5 mM fructose, or transfected with miR-377 mimic/inhibitor and TXNIP siRNA, or co-incubated with p38 MAPK inhibitor, demonstrating that miR-377 overexpression activates the O 2 − /p38 MAPK/TXNIP/NLRP3 inflammasome pathway to promote oxidative stress and inflammation in fructose-induced podocyte injury. Antioxidants pterostilbene and allopurinol were found to ameliorate fructose-induced hyperuricemia, podocyte injury, and albuminuria in rats. More importantly, pterostilbene and allopurinol inhibited podocyte miR-377 overexpression to increase SOD1 and SOD2 levels and suppress the O 2 − /p38 MAPK/TXNIP/NLRP3 inflammasome pathway activation in vivo and in vitro, consistent with the reduction of oxidative stress and inflammation. These findings suggest that miR-377 plays an important role in glomerular podocyte oxidative stress, inflammation, and injury driven by high fructose. Inhibition of miR-377 by antioxidants may be a promising therapeutic strategy for the prevention of metabolic syndrome-associated glomerular podocyte injury. [ABSTRACT FROM AUTHOR]

Clear cell renal cell carcinoma (ccRCC) is the most common form of neoplasm occurring in the adult kidney. Although significant advances have been made in treatment for ccRCC, a significant percent of patients have no benefit from currently available treatment option. MicroRNAs (miRNAs) have been reported to play central roles in regulating tumor progression, and are being explored as potentially more effective therapies and diagnostic tools for various cancers. The transcription factor E26 transformation specific-1 (ETS1) is believed to be intimately involved in tumor progression, and is frequently upregulated in tumors, including ccRCC. However, few studies have investigated the implications of ETS1 in ccRCC, and no studies have investigated the dysregulated mechanisms responsible for aberrant ETS1 expression in ccRCC. We used databases, clinical samples and target prediction algorithms to investigate aberrant miR-377 expression and potential targets in ccRCC. Our results indicate that miR-377 is downregulated in ccRCC, and that miR-377 can regulate the expression of ETS1. Further, using cell cycle analysis, MTT, luciferase and knockdown experiments, we found evidence to suggest that ETS1 is central in the development of the proliferative, metastatic and invasive phenotype of ccRCC cells. Furthermore, miR-377 was found to directly downregulate the expression of ETS1 and reduce the ability of ccRCC cells to proliferate, migrate and invade. As miR-377 was found to be differentially expressed in ccRCC, and in light of the apparent central role of ETS1 in tumor development, our results indicate that miR-377 could be useful for ccRCC diagnostics, prognostics and therapeutics. [ABSTRACT FROM AUTHOR]