Cloning And Sequence Analysis Of Light-Heavy Chain Variable Region Gene Of Anti-ER-?36 Monoclonal

Breast cancer is one of the most common malignancies in women.The incidence of breast cancer in the world has increased year by year.In developed countries such as Europe and the United States,breast cancer has become one of the major causes of death in women.In many large and medium cities in China,breast cancer has also been the highest incidence of female cancers.It is a serious threat to women's health.Current comprehensive treatment of breast cancer includes surgical excision,medical treatment and local radiotherapy.In the field of medical treatment,breast cancer is divided into different subtypes and the corresponding treatment.Triple negative breast cancer(TNBC)refers to a special type of breast cancer that ER,PR and Her-2 are all negative,is associated with early recurrence,rapid progress and poor prognosis.Due to the lack of corresponding receptors,routine endocrine therapy and targeted Her-2 treatment are ineffective.Our previous work revealed that ER-a36 expression was significantly higher in triple-negative breast cancer.Downregulation of ER-a36 cell line MDA-MB-231 resulted in more sensitivity to the chemotherapeutic drug paclitaxel,their migration and invasive ability were significantly reduced,and these changes were not related to the presence of estrogen.These results suggest that ER-?36 may also be a potential target for the treatment of triple-negative breast cancer.Because ER-a36 is mainly expressed in the cell membrane and cytoplasm,specific monoclonal antibody may have a neutralization effect and thus play their biological function.If we can further reconstruct the ER-a36 monoclonal antibody,we may improve its biological effect and clinical application value.In this experiment,we used the hybridoma cells that secreting the specific anti-ER-a36 monoclonal antibody.Total RNA was extracted from the cell,and the light and heavy chain variable region gene of the monoclonal antibody was amplified by RT-PCR and then sequenced.The results showed that the heavy chain variable region gene 438 bp belonged to the mouse monoclonal antibody IgM heavy chain.The light chain variable region gene of the antibody is 416 bp,which belongs to the light chain of immunoglobulin,the variable region gene of ? chain.The cloning and sequence analysis of the variable region gene is a key step of the genetic engineering reconstruction of the monoclonal antibody.