Analysis Of The Effect Of Delta S2 SF-1a PRLR On The Transcriptome Of Human Breast Cancer Cells MCF-7 By RNA-seq Technique

Prolactin(PRL)is a single chain polypeptide hormone secreted by the pituitary gland and some pituitary tissue,such as the mammary gland.PRL plays an important role in water and electrolyte balance,development and growth and other physiological activities.PRL binds to PRLR on the surface of the target cell membrane to initiate the corresponding intracellular signal transduction,allowing PRL to achieve its biological function.?S2 PRLR is a variant of PRLR found by Tan et al in human breast cancer cells as well as prostate cancer cells,cloned and reported for the first time.Its main structural features are: the outer membrane S2 subdomain of the receptor(membrane outer region contains S1,S2 two domains)is missing.The PRLR variants currently cloned by Tan are ?S2 LF-PRLP,?S2 SF1a-PRLP,?S2 SF1b-PRLP.A large number of experiments show that overexpression of ?S2 LF can significantly promote the proliferation of breast cancer cells.In addition,Zhang has performed a transcriptome analysis of gene expression in ?S2 LF-PRLR in breast cancer cells(MCF-7)during its graduate studies.And found 30 differentially expressed oncogene,27 tumor-associated genes were only expressed in ?S2 LF-PRLR overexpressing MCF-7 cells.These results suggest that ?S2 PRLR plays an important role in the development and progression of human breast cancer.At present,it is clear that LF-PRLR activates the JAK-STAT pathway and SF1 b plays a negative role in regulating LF.However,there was no consensus on the effect of SF1a-PRLR.To further investigate the effects of SF1a-PRLR and ?S2 SF1a-PRLR,it is important that total transcriptase analysis was performed on SF1a-PRLR and ?S2 SF1a-PRLR overexpressing MCF-7 cells and analysis the differential expression genes.We transfected human breast cancer cells(MCF-7)with SF1a-PRLR(group G8)and?S2 SF1a-PRLR(group 1a)by virus-mediated gene transfer.The control group(Con group)was transfected with an empty virus that did not carry any exogenous gene.Cells were incubated for 48 hours and the total RNA was extracted for RNA sequencing(RNA-Seq).The data obtained after sequencing were uploaded to the website of the University of California,Santa Cruz Genome Browser(genome.ucsc.edu).The results showed that: 1)In the blank control group and overexpression SF1a-PRLR genome,4862 gene was differentially expressed,wherein the decreased expression genes 2175,increased expression of genes 2687;In the blank control group and overexpressing ?S2 SF1a-PRLR genome,4905 was differentially expressed,wherein the decreased expression genes 2140,increased expression of genes 2765 genes.In the overexpression of ? S2 SF1a-PRLR and overexpressing the SF1a-PRLR gene,11 genes were differentially expressed.2)KEGG pathway enrichment analysis was performed on differential gene expressing between the blank control group and the SF1a-PRLR overexpressing group.It was found that a variety of signaling pathways involved in the regulation of tumor growth and maintenance of tumor cell viability were enriched,such as MAPK signaling pathway,TNF signaling pathway,TGF-beta signaling pathway,PI3K-Akt signaling pathway,cAMP signaling pathway,Hedgehog signaling pathway,Hippo signaling pathway.3)2)KEGG pathway enrichment analysis was performed on differential gene expressing between the blank control group and the ?S2 SF1a-PRLR overexpressing group.A number of signaling pathways involved in the regulation of tumor growth were found to be enriched,such as MAPK signaling pathway,TNF signaling pathway,TGF-beta signaling pathway,PI3K-Akt signaling pathway,ErbB signaling pathway,Hedgehog signaling pathway,Hippo signaling pathway.According to the results of this experiment,the following conclusions are drawn: The expression of SF1a-PRLR gene and ?S2 SF1a-PRLR gene could affect the gene expression of MCF-7 breast cancer cells.2)Overexpression of SF1a-PRLR gene and ?S2 SF1a-PRLR Genes can influence cell proliferation through MAPK signaling pathway,TGF-beta signaling pathway,PI3K-Akt signaling pathway,Hedgehog signaling pathway and other signaling pathways.This study is of great significance for understanding the molecular mechanism of SF1a-PRLR and ? S2 SF1a-PRLR in human breast cancer and laying a solid foundation for further exploring the effect of SF1a-PRLR and ?S2 SF1a-PRLR.