Construction And Preliminary Identification Of ULK1 Transgenic Mice

Prostate cancer?PCa?is one of the most commonly diagnosed malignant tumor in the western world.Although it usually occurs in men older than 60,there appeared to be more and more young patients suffering from prostate cancer in recent years.In China,the frequency of prostate cancer has been increased due to the expanding of aging population and the transforming of dietary structure.Androgen deprivation is frequently-used in prostate cancer therapy,but it will lost its efficacy gradually after a period of treatment because the cancer has developed into castration resistant prostate cancer?CRPC?which is insensitive to androgen?Autophagy is an evolutionarily conserved cellularcomponents degradationprocess and it plays an important role for cell survival and development.In cancer cells,autophagy is a double-edged sword,as it can either suppress tumor or provide energy for cancer cells.ULK1 is a Ser/Thr kinase,and it is quite essential to autophagy initiation in response to energy deficiency or nutrition deprivation.By studying about the function of ULK1 in prostate cancer initiation and developing process,we can make further research into prostate cancer mechanism in the new aspect of autophagy.Animal models have been an important tool in biology research.Prostate cancer mice model have been widely used in mechanism research and drug screening.Traditional prostate cancer mice models is made by transplanting cancer cells or tissue isolated from primary cancer or metastasis tumor into mice.With the development of genetic engineering techniques,we can get transgenic mice by altering the mice genome,it enable us to evaluate relevant target gene and associated signaling in prostate cancer.In this study,molecular biology techniques are used to construct ULK1 transgene plasmid.After ULK1 transgenic mice is obtained,it is intercrossed with PB-Cre mice to generate ULK1;PB-Cre transgenic mice,which is used to investigate whether ULK1 could influence prostate cancer initiation.First,mice genotype was characterized by PCR genotyping and immunohistochemistry?IHC?,then histopathology analysis is made by H&E staining the prostate tissue which get from transgenic mice and its littermate control by anatomical analysis?Results show that ULK1 cannot initiate prostate cancer during the first three month.In order to study the function of ULK1 in the prostate cancer development,ULK1;Pten^f/f;PB-Cre transgenic mice are constructed.Tissue taken from 4-5M and 5-6M transgenic mice and Pten^f/f;PB-Cre mice in the same age are stained by H&E,and histomorphology is compared.From the tissue slice,it can clearly observed that prostate is developing in a range of steps from hyperplasia to low-grade PIN?prostatic intraepithelial neoplasia?,then grew into high-grade PIN,which is coincidence with the development pattern of prostate cancer.At the same age level,ULK1;Pten^f/f;PB-Cre mice prostate develop in a higher speed in contrast with Pten^f/f;PB-Cre mice.So it can be preliminarily determined that ULK1 can promote prostate cancer development.In summary,ULK1 can accelerate the process of prostate cancer to some extent,suggesting it can become a potential prostate cancer treatment target,thus achieve effective control of cancer process through genetic method.