Study On The Function Of Long - Chain Non - Coding RNA CACNA1G - AS1 In Keloid

Background:Keloid is a kind of benign proliferative diseases. Because of its invasion to the surrounding normal tissues, it can cause dysfunction and aesthetic deformity with frequent pruritus and pain, making patients suffer physically and psychologically. Treatment of keloids still lack effective methods, which makes it a difficult problem of plastic surgeon. Long non-coding RNA, namely lncRNA, refers to a class of non-coding RNA that length longer than 200bp. LncRNA can regulate cellular processes such as cell cycle and proliferation, differentiation, apoptosis in the level of transcription, post-transcription, epigenetics and so on. Some lncRNA have been found to involved in the development and progression of malignant tumors and skin disease. In our preliminary study, we first in the world found the expression of CACNA1G-AS1, namely CAS1, is specificly high in keloid tissue, which suggest it may be involved in the formation mechanism of keloid.Objective:To explore the biological function in human keloid fibroblasts such as cell proliferation, migration, cytokine secretion, collagen secretion, and control of expression of calcium channel proteins.Methods:We collect 3 samples of keloid from patients received treatment in Peking Union Medical College Hospital, who have signed informed consent form. After cell cultured, we use siRNA to interfere CAS 1 expression, examining the efficiency of transfection and interference. Then use MTS assay and c-flow test to see the influence on cell proliferation, use scratch experiments to see the influence on cell migration, use qRT-PCR to see the influence on secretion of cell factor, collagen, and expression of calcium channel protein. Every experiment was compared to the control group which transfected with negative control sequence.Results:Compared with negative control group, the inference efficiency has reached 50%. The expression of CACNA1G and type Ⅰ collagen have significantly up-regulated in experimental group, while the expression of TGF-beta, type Ⅲ collagen haven’t show any difference. The scratch experiment shows a longer healing time of experimental group, while there is no difference between control group and experimental group in cell proliferation.Conclusions:CAS1 may promote the expression of calcium channel protein CACNA1G and type Ⅰ collagen, also has a positive effect on cell migration in human keloid fibroblast, making it a potential target of novel therapy.