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[Abstract]:Prolactin receptor (PRLR) is one of the members of the type I cytokine receptor superfamily and is a single transmembrane receptor. PRLR plays a key role in the biological effects of prolactin (PRL). Under the effect of PRL, PRLR is dimerized, thus initiating cell signal transduction and achieving its biological effect. The preS2 LF-PRLR is a variant of a PRLR that is cloned and first reported by Tan et al. in human breast cancer cells. The main structural feature is that the subdomain of the extracellular domain S2 of the receptor is deleted (consisting of 100 amino acids and the other subdomain is S1). It has been shown that, in the absence of PRL, the TS2 LF-PRLR can also dimerize, that is, the so-called self-dimerization, and to produce biological effects such as promoting cell proliferation and the like. It is suggested that SS2 LF-PRLR may have important molecular and pathological significance in the occurrence of human breast cancer. After the activation of the PRLR, the expression of the corresponding gene is adjusted by activating the JAK2-STAT5 cell signal transduction pathway downstream of it, which in turn results in phosphorylation, dimerization and nuclear transfer of the STAT5, thereby binding to the promoter region of its target gene. If the S2 LF-PRLR is lost due to the S2 substructure outside its cell, we speculate that this deletion has changed the molecular nature of the PRLR, thus affecting the cell transduction pathway and then changing the expression profile and the expression level of the target gene, and finally changing the biological behavior of the breast cancer cell, For example, excessive proliferation of cells, and the like. In this study, we used the virus-mediated gene transfer method to transfect the human breast cancer cell (MCF-7) with the adenovirus carrying LF-PRLR (LF-PRLR group) or the wild-type S2 LF-PRLR (FS2 LF-PRLR group) c DNA, and the control group (Con) was transfected with an empty virus that did not carry any foreign gene. After the cells continue to be cultured for 48 hours, the total RNA of the cells is extracted and then the RNA-Seq is performed and the sequencing data is uploaded to the genome browser website (genome.ucsc.edu) of the University of California, Santa Cruz. The results showed that: (1) After the overexpression of VS2 LF-PRLR, the expression of the gene was larger (that is, the difference between the three groups of Con, LF-PRPR and FS2LF-PRLR was large), mainly in the chromosome 11,12,14,16,17,19,20, while the difference in the chromosome 4,13,18,21 was relatively small; (2) the gene affected by it not only had the coding gene, The non-coding gene is also included. The number of expression genes (5% of the lowest RPKM value) of the three groups of Con, LF-PRPR and FES2 LF-PRLR were 19648 (coding gene 17594 non-coding gene 2054),20247 (coding gene 18093, non-coding gene 2154) and 19500 (encoding gene 17472, non-coding gene 2028), respectively. (3)192 genes in the whole transcriptome were expressed only in the FES2 LF-PRLR MCF-7 cells (in which there were 118 of the coding genes and 74 non-coding genes), and the 483 genes were not expressed in the FS2 LF-PRLR MCF-7 cells, while in the LF-PRLR and Con group (where the coding gene had 362 and the non-coding gene was 121); The 18924 genes were expressed in three groups (1753 of the coding genes and 1871 non-coding genes), and there were 3 (all non-coding genes) of the genes expressed in the SS2LF-PRLR group. In the three groups, there were 1 (also non-coding gene) of the low-expression genes in the FS2 LF-PRLR group, and (4) the overexpressing of the FS2 LF-PRLR resulted in the change of the expression of the tumor-related genes. There were 30 tumor-related genes in the transcriptome of three groups, and 27 tumor-related genes were expressed only in the QS2 LF-PRLR group, but not in the Con group and the LF-PRLR group. (5) The expression level of the apoptosis-related gene was altered by the overexpression of VS2 LF-PRLR. There were differences in the expression of 11 apoptosis-related genes in the three groups, of which 9 of the apoptotic genes were expressed in both the Con and LF-PRLR groups, but the expression of the two anti-apoptotic genes in both the Con and LF-PRLR groups was not expressed. (6) Partial microRNA (micro-RNA) is also the target of the action of TS2 LF-PRLR. There were 15 microRNAs in the transcriptome of three groups, and 8 Micro RNAs were expressed only in the FS2LF-PRLR group and the 4 Micro RNAs were not expressed in the other two groups. (7) The PS2LF-PRLR has the function of regulating the expression of sno RNA, and there are 19 difference in the expression of 19 sno RNA in the three groups,13 of which are box C/ D sno RNAs and 6 are box H/ ACA sno RNAs. Of these,10 of the sno RNA genes were not expressed in the other two groups, while the other two groups were not expressed in the other two groups, while the other two groups were not expressed. According to the results of the experiment, the main conclusions of this study are as follows: (1) The overexpression of VS2 LF-PRLR can affect the expression of human breast cancer cells (MCF-7). It is affected by the coding gene and the non-coding gene. (2) The effect of the transcription of the human breast cancer cell (MCF) gene is clearly different from that of the full-length receptor LF-PRLR. (3) The expression of TS2 LF-PRLR in the control of tumor-related genes affects the biological behavior of breast cancer cells, such as cell proliferation, and so on. (4) The expression of the apoptosis gene is down-regulated by the anti-S2 LF-PRLR, thereby promoting the proliferation of the breast cancer cells. (5) The expression of the micro-RNA (Micro-RNAs) is affected by the FS2 LF-PRLR. (6) The expression of sno RNA was regulated by the control of SS2 LF-PRLR. This study is of great significance to understand the molecular mechanism of TS2 PRLR in human breast cancer.
【學(xué)位授予單位】：吉首大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R737.9;Q78

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