Expression Of Med-19 In Cutaneous Malignant Melanoma And Its Effects On The Biological Behavior Of A375 Cells

Background and Objectives Malignant melanoma(MM)is a highly malignant tumor derived from neural crest melanocytes,common in skin and the mucosa of the organs,stays hidden,but occurs in the early stage as local and distant metastasis.Disease progression is rapid and highly malignant with median survival rate of only 7.5 months.The prognosis is poor and the fatality rate is high.In the past few decades,incidence is rising and is the leading cause of mortality by skin diseases.According to the epidemiological survey,in recent years,the incidence and death rate of China’s Cutaneous Malignant Melanoma(CMM)are an upward trend.Currently available treatment is mainly by surgical resection,chemotherapy combined with radiotherapy after surgery,molecular targeted therapy to improve the survival of patients with malignant melanoma.Lack of definitive pathogenesis makes it difficult for early diagnosis and management.It is important to explore the mechanism of the occurrence and development of malignant melanoma,searching for new tumor markers and therapeutic targets,.Furthermore,improve the early diagnosis and treatment of malignant melanoma,monitor the recurrence and metastasis.Mediator complexes(Med)is a macromolecular complex composed of multiple protein subunits,which have been confirmed to promote activation,enhance basal transcription,promote carboxy terminal domain(CTD)cell transcription(polymerase subunit hydroxyl terminal domain phosphorylation of CTD)function in the regulation of cell growth.Each processes differentiation and apoptosis directly or indirectly play a role.Mediator complex subunit 19(Mediator 19 or Med19)is one of its subunits,and is located in the 11q12.1 position in the human chromosome,encoding gene called LCMR1(lung cancer metastasis related protein,Geb Bank No.AAN16075).Previous research shows that the expression of Med19 in lung cancer,breast cancer,gastric cancer,liver cancer and other organs is related to the formation and development of the tumors.Changing the expression of a protein in a cell is a common research idea that studies the role of the protein in cells.Previous studies on the malignant biological behavior of Med19 genes,such as proliferation,apoptosis,differentiation,and invasion,provide a new therapeutic consideration for gene therapy of tumors.RNA interference(RNAi)refers to the phenomenon that the homologous m RNA is highly degraded by double stranded RNA(ds RNA),which is highly conserved in the evolution process.RNAi technology can be used to specifically remove or close the expression of specific genes,was widely used to explore gene function,infectious diseases,malignant tumor treatment.There are a lot of studies at home and abroad have proved that the use of RNAi silencing Med19 gene inhibition of human breast cancer cells.Human lung cancer A549 cells and other tumor cells to detect the interference after its proliferation,apoptosis,invasion,metastasis and other effects for these malignant tumors,treatment provides new treatment ideas.And on the Med19 and skin melanoma related research,there is no relevant reports.Therefore,the aim of this study is to detect the expression of Med19 in normal tissues of skin melanoma and peritumoral tissues by immunohistochemical method,and to study the correlation between the expression level and the malignant phenotype of melanoma.The expression of melanin,the expression of Med19 m RNA in A375 cells was detected by RT-PCR,cell proliforation was detected by MTT assay.Apoptosis was detected by flow cytometry.To investigate the correlation between Med19 and malignant melanoma.Part one The expression and significance of Mediator complex subunit 19 in cutaneous malignant melanomaResearch Purpose:To study the expression of Med19 protein in normal tissues of skin melanoma and normal tissues,and to analyze the correlation between Med19 protein and clinical parameters.Research method:1.Immunohistochemical method was used to detect the expression of Med19 protein in normal melanoma tissues and normal tissues.2.Statistical analysis of the expression of Med19 protein and the clinical parameters of patients were analyzedResults:1.The expression of Med19 protein in 52 cases of melanoma and 20 cases of normal tissues was detected by immunohistochemistry.The results showed that the expression of Med19 positive protein in 52 cases of melanoma was 86.54%(45 cases)and 20 cases The positive expression of Med19 protein was 15.0%(3 cases),the difference was statistically significant(P <0.05).2.The expression of Med19 protein in the skin melanoma tissue was higher than that in the normal tissues of the cancerous tissues,and the expression of Med19 in the tissues with late lymph node metastasis and TNM stage was higher than that in the untreated patients with TNM stage(P <0.05).(P> 0.05).Conclusion: There is no significant correlation between age,sex and primary ulcer.The expression level in malignant melanoma tissues was higher than that in normal tissues(P <0.05).Conclusion:1.The expression of Med19 protein in skin melanoma tissue was significantly up-regulated compared with normal tissue,suggesting that Med19 may be involved in the development of melanoma.2.Med19 in the expression of melanoma in the skin in patients with lymph node metastasis,TNM staging in patients with higher than those without metastasis,TNM staging earlier patients(P<0.05),differences were statistically significant.These results suggest that Med19 may be related to the biological behaviour of malignant melanoma.Part two The expression of Mediator complex subunit19 on the proliferation and its effect on the metastasis of the A375 cells of malignant melanoma.Research Purpose:In the first part of the experiment,Med19 was observed in malignant melanoma tissue.Normal cancer tissue were expressed,and the skin melanoma tissue Med19 protein expression was significantly higher than the normal tissue weeks.Statistical analysis confirmed the expression of lymph node metastasis,TNM staging was positively correlated with sex,age,etc.,suggesting that Med19 may be associated with the biological behavior of melanoma.The expression of Med19 m RNA and Med19 protein was detected by RT-PCR.The expression of Med19 gene on the proliferation and apoptosis of melanoma A375 cells was observed by MTT and apoptosis test.To observe the effect of silencing Med19 gene on the proliferation and apoptosis of malanoma A375 cells.Research Purpose:By si RNA silence Med19 expression,the effects on the proliferation and metastasis of malignant melanoma A375 cells were observed.Research Method:1.The expression of Med19 m RNA in melanoma A375 cells was detected by RT-PCR.2.The effect of Med19 silencing on the proliferation of melanoma A375 cells was detected by proliferation assay.3.The effect of Med19 silencing on the apoptosis of melanoma A375 cells was detected by flow cytometry.Results:1.A375 cells si RNA expression can inhibit the expression of Med19 gene m RNA.2.Med19 gene silencing has a significant inhibitory effect on the proliferation of melanoma A375 cells.3.Med19 gene silencing can cause significant apoptosis of melanoma A375 cells.Conclusion:Si RNA can effectively silence the expression of Med19 gene in melanoma A375 cells.Silencing Med19 gene can induce the apoptosis of A375 tumor cells and inhibit the growth of A375 cells.The results show that Med19 is involved in the development and development of melanoma,and it is expected to become melanin tumor clinical diagnosis,an important basis for staging,and can be used as a new target for cancer treatment.