Investigation About The Influence Of Rab34 On The Development Of Zebrafish And The Migration Or Invasion Of Breast Cancer Cells

Breast cancer is one of the higher incidence of female malignant tumors,which is accounts for about 7-10%of the all kinds of malignant tumors.However,there are no specific diagnosis biomarkers or therapeutic target for breast cancer,our lab found that Rab34 which is small molecule GTP apoenzyme.is highly expressed in breast cancer patients of high degree and whose tissue is adjacent to carcinoma.Because the embryonic development and the deterioration of tumor is often accompanied by the epithelial mesenchymal transdifferentiation,thus,our main task is to study the correlation between the Rab34 and EMT,further to investigate the influence of Rab34 on embryonic development and the invasion or migration of breast cancer cells and finally to look for specific diagnosis biomarkers or therapeutic target for breast cancer.Zebrafish is our primary study object to explore the influence of Rab34 on zebrafish development.Firstly,there are zRab34a and zRab34b in zebrafish is different from the human,the test of immunofluorescence found that zRab34a is mainly located in the Golgi and can raise lysosome to the around the nucleus,which is same with hRab34.However,there is no the phenomenon about zRab34b.Next,we build zRab34a overexpression model and knockout zRab34a model using CRISPR/cas9 gene-editing techniques.Moreover,we screen the homozygous strain.Taking advantage of whole-mount in situ hybridization,we found Whether overexpression of zRab34a or knockout zRab34a could affect the growth of zebrafish and the group of knockout zRab34a Significantly inhibit the development of zebrafish.Furthermore,we put zebrafish and nude as our study object to do tumor cell transplantation in vivo experiment exploring the influence of Rab34 on breast cancer cell migration/invasion ability.Finaly,We found that knockdown Rab34 could inhabit the breast cancer cell migration or invasion ability.For further research that whether there is a correlation between Rab34 and EMT,we selected E-cadherin which is one of the EMT common markers at embryos early developmental stage of zebrafish.we preliminary found that Rab34 can promote the occurrence of EMT.Our studies have shown that Rab34 may be promote the occurrence of EMT and further affect the development of zebrafish embryo and the breast cancer cell migration or invasive ability,in the near future,Rab34 may could as a potential diagnostic marker or drug therapeutic target to breast cancer.