The Expression And Function Of HSDL2 In Human Malignant Melanoma

Objective: Malignant melanoma(malignant melanoma,MM)is a malignant tumor transformed by melanocytes.Melanocytes are differentiated from the neural crest,so MM occurs in tissue or organs that are differentiated from neural crest cells,such as oral cavity,nasal cavity,gastrointestinal mucosa,cranium and other parts。But the highest incidence of MM in the organ is skin.The incidence of malignant melanoma is different from the different human species and the different ultraviolet intensity in the living area.Melanin is produced by pigment cells and is used to protect the skin from UV damage.Data show that the incidence rate of MM in Caucasians was significantly higher than that in African and Asians.The incidence of MM in the area of the same skin color with strong ultraviolet radiation is high.Irritation and incorrect treatment of pigmented nevus are also the cause of MM.Our people’s knowledge of malignant melanoma is generally not high.Long term outdoor workers have no effective self protection to reduce skin damage by ultraviolet light Long term outdoor workers have no effective self-protection to reduce skin damage by ultraviolet light,especially for pigmented nevus,such as plantar,lumbar,oral mucosa and other vulnerable locations.In addition,the improper treatment of pigmented nevus,such as repeated friction,chemical drugs,freezing,and no thorough excision,which can induce the malignant change of pigmented nevus.When a patient was found to have skin ulceration or swelling,he would go to the hospital,but at this time,malignant melanoma often had infiltration of surrounding tissues and lymph node metastasis,and lost the best time of surgical treatment.The data showed that the 5 year survival rate of the 0 phase MM was97%,and the 5 year survival rate of the IV period MM was only 10%.The incidence of MM in China is not high,it accounts for about 4%~5% of systemic malignancies,but the mortality rate ranks the first among all skin tumors,and the 5 year survival rate of MM in China is less than 10%.There is no effective screening for early malignant melanoma,and this is one of the reasons for the poor survival rate of MM patients.So studying on malignant melanoma cells from a genetic perspectiveand finding effective biological targets,are important methods to improve diagnosis rate of the early malignant melanoma.Meanwhile,effective biological targets can also provide new clinical ideas for the treatment and prognosis of MM.Methods:The high intension gene function screening platform is used to screen the new functional positive gene HSDL2 in melanoma cells.Construcing different targeting RNA lentiviral vector infection1 to the HSDL2 gene using Western Blot extraneous to validation the target effectiveness and screening effective targets,in the tool cell293 T.1.The q PCR method was used to detect the high richness of the HSDL2 gene in different tool cells of m RNA.2.The design of RNA jamming target and double stranded DNA oligo are designed with the HSDL2 based template,and construct lentivirus vector.3.In vivo,A375(human malignant melanoma cells)was knocked out of the tumor cell HSDL2 gene,and the A375 stable cell bead of the HSDL2 defect was constructed after screening.4.Detection of gene knockout rate of HSDL2 gene at m RNA level by q PCR.5.Sh HSDL2(A375 cell and HSDL2 gene sh RNA virus infected cell group)were detected by Celigo cell count detecting cell growth,MTT detection,PI-FACS cell cycle detection and Annexin V-APC single staining method.Results:After sh RNA lentivirus infection,the expression level of HSDL2 gene in A375 cells was significantly inhibited by RT-PCR method(p<0.05),and the knockdown efficiency was 90.8%.2.The detectiong of cell growth showed that the proliferation of sh HSDL2 group(normal A375 cells plus HSDL2 gene sh RNA virus infected cell group)was significantly inhibited compared with sh Ctrl group(normal A375 cells,plus negative control virus infected cells).3.The results of MTT detection showed that the proliferation of cells in group sh HSDL2 was slower than that in group sh Ctrl.4.Cell cycle detection showed that the cell of sh HSDL2 group was increasing in S stage(P<0.05),increasing in G1 phase(P<0.05),and decreasing in G2/M stage(P<0.05).5.Apoptosis detection: compared with the sh Ctrl group,the number of apoptotic cells in group sh HSDL2 increased(P<0.05).Conclusion: Interfering target gene HSDL2 can effectively reduce melanoma cell A375 cell activity,inhibit cell proliferation and promote cell apoptosis,which may be an effective molecular biological target of MM,and provide new ideas and methods for the diagnosis,treatment and prognosis of MM.