The Effect Of MicroRNA-152 On The Proliferation And Collagen Synthesis Of The Keloid Fibroblast

Background:Keloids are bebign tumors of the skin, which are caused by exceedingly collagen synthesis in the damage healing process, characterized by overgrowth of lesions, forming red, stiff lumps beyond the original boundarys. Keloids are also accompanied by itching, pain, infection, dysfunction, and even cancer as well as harmful to one’s appearance. It is one of the most intractable problems for plastic surgeons. So far there is not a certain effective treatment of keloids for clinical doctors. Keloids also have some characteristics like tumors. The formation of keloids is regulated by the multiple genes, cell factors and immune factors.MicroRNAs are small non-coding RNA molecules, containing about 22-25 nucleotides.They have important regulation function in eukaryotic gene transcription level and/or translation level. Since it’s first discovered in 1990s, microRNA has became the hightlight for researchers. Aberrant expression of microRNAs has been implicated in numerous diseases, such as cancer, myocardiac disease, skin fibrosis and metabolism desease. It has been demonstrated that there are multiple roles for microRNAs in these biological processes.MicroRNA-based therapies are under investigation. Former studys have found that a variety of microRNAs have aberrant expression in keloid fibroblasts, including microRNA-152, but its role in the occurrence and development of keloid remains unclear.Objects:To investigate the proliferation and collagen synthesis of cultured human keloid fibroblast after transfecting with microRNA-152 mimics in vitro. Discussing the possible mechanisms, and provide the basis for the prevention and treatment of keloid.Methods:Collecting fresh keloids specimens and culture keloid fibroblasts in vitro, using liposome 2000 transfect the keloid fibroblasts by microRNA-152 mimics, using real-time fluorescent quantitative PCR technique to detect the microRNA-152 expression level, CCK method to detect cell apoptosis, hydroxyproline colorimetry analysis the total collagen synthesis of the supernatant fluid; western blot to detect the collagen protein expression in the cell; Real-time PCR to detect the mRNA levels of COL1A,COL3A1 and TGF beta 1 gene in fibroblast cells.Results:In our research, the microRNA-152 mimics are successfully transfected into KF. Comparing with the control group, the quantity of cell apoptosis in the microRNA-152 overexpress group is reduced, OD value at 48h (0.699±0.031) vs. (0.588±0.0071), p <0.05; 72h (0.726±0.0095) vs. (0.633±0.011), p<0.05; Colorimetry analysis found that the hydroxyproline content (μg/ml) in supernatant fluid is increased(3.909±0.015) vs.(2.311±0.075), p<0.05; the expression of Type I and Type III collagen in KF cells increased separately; Real-time PCR detection found that the mRNA levels of Type I, Type Ⅲ collagens and TGF beta 1 gene in the fibroblasts increased separately (1.75±0.019) vs.(1.063±0.017), p<0.05; (1.356±0.074) vs.(1.045±0.061), p< 0.05; (2.12±0.03) vs. (1.08±0.033), p<0.05.Conclusion:MicroRNA-152 may promote the keloid fibroblast proliferation and collagen synthesis. Therefore microRNA-152 may be one of the important molecules in the pathogenesis of keloid.MicroRNA-152 is expected to become a new biomarker and a potential target for the treatment of keloid.