MicroRNA-449a Regulates The Expression Of Bcl-2 In HaCaT Cells

Currently the most studied microRNAs(miRNAs),which are highly conserved non-coding RNA molecules composed of 19-25 nucleotides,play a key role in apoptosis,cell cycle arrest,and cell differentiation.In this experiment,we mainly studied MicroRNA-449 a,which is a potent inducer of apoptosis,cell cycle arrest,and cell differentiation,Therefore,we further explore the role of miR-449 a in the regulation of apoptosis and provide guidance for the future treatment and prognosis of tumors.The Bcl-2 family is a mitochondrial-associated protein that regulates apoptosis and was cloned from lymphoma by Tsujimoto et al.and named B-cell lymphoma/leukemia-2(Bcl-2).Bcl-2 family members can be divided into two major categories: 1 anti-apoptotic protein types: mainly including Bcl-2,Bcl-xL,Bcl-w,A1,Mcl-1,etc.,have anti-apoptotic effect.2 promote the apoptosis of proteins: This type of member has a role in promoting apoptosis,including: Bax,Bak1,Bok,Bid,Bad and so on.The Bcl-2 family protein is an essential apoptotic regulator and is encoded by the BCL2 gene located on chromosome 18q21.3.As a type of apoptosis-related gene,BCL-2 plays an important role in regulating apoptosis,reducing ischemia-reperfusion injury and anti-tumor,and is also a key factor in the occurrence and development of cancer.In order to further study the expression and regulation mechanism of BCL-2,it is of great significance to better treat cancer and other diseases.Objective:In this study we investigated the role of miR-449 a in the regulation of Bcl-2 expression in human immortalized keratinocytes(HaCaT),as well as the mechanisms that induce apoptosis,and further guide the treatment and prognosis of tumors.Materials and Methods: First,HaCaT cells were cultured and transfected into HaCaT cells.Corresponding RNA extraction and protein extraction were performed on HaCaT cells at 24 h,48h and 72 h after transfection,respectively.The samples were frozen at minus 80 degrees Save the refrigerator.The extracted RNA and protein were detected by reverse transcription polymerase chain reaction(RT-PCR)and Western blot(Western blot)method to detect the expression of bcl-2.Results: Transfection of miR-449 a HaCaT cell line,RT-PCR and Western blot detection methods and found that the transfected miR-449 a longer BCL-2 expression was significantly decreased.Conclusion: miR-449 a is a potent inducer of apoptosis,which can effectively inhibit the expression of anti-apoptotic gene BCL-2 and thus may promote apoptosis.