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The Role of MicroRNAs and Target Gene in Uveal Melanoma and Gastric Cancer Development and Progression

Author: SunQingMin
Tutor: WangBin
School: Nanjing Medical University
Course: Pharmacology
Keywords: MicroRNA-27a microRNA-9 genistein ZBTB10 Uveal melanoma Gastric cancer Polymorphism
CLC: R735.2
Type: Master's thesis
Year: 2009
Downloads: 64
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Abstract

MicroRNAs (miRNAs) are found in recent years, a class of endogenous non-coding the small molecules RNAs, through direct degradation of target gene mRNA or inhibiting their translation, thus the negative regulation of post-transcriptional gene expression. Despite its potential biological function is not yet fully elucidated, has found that miRNAs may be involved in the regulation of cellular processes including apoptosis, proliferation, and differentiation. Studies have shown that some of the miRNA can be used as tumor diagnostic markers or therapeutic targets including colon cancer, breast cancer, lung cancer, gastric cancer, etc.. Recent studies have shown that the abnormal expression of miRNAs can also be used as a transfer of risk of uveal melanoma markers. According to national health statistics department statistics, in the cities of cancer is the primary cause of death, and is ranked third in the countryside. Therefore, to improve the awareness and understanding of the molecular mechanism of cancer is particularly important to promote the development of new therapeutic strategies. In the present study, we observed that the role and significance of miRNAs in the adult eye the highest incidence of the primary tumors of uveal melanoma and common tumor incidence of gastric cancer development. Role and microRNAs mechanism Genistein first part of Genistein on human uveal melanoma cells as an isolated soybean isoflavones, have been found to affect a range of cellular processes or enzymes, such as the cell cycle, apoptosis, angiogenesis play The anti-cancer effects. However, as far as we know, there is no contact between the study focuses on miRNAs mediated transcriptional regulation and genistein anticancer effects. MiR-27a as a proto miRNA expression in a variety of cancer cells, and previous studies have shown that in breast cancer cells transfected with antisense miR-27a can significantly increase the expression of ZBTB10 (Zinc finger and BTB domain containing 10). MiR-9 was found to be associated with tumor invasion and metastasis. Therefore, we investigate the role of genistein uveal melanoma and observe the expression of miR-27a and its target gene ZBTB10. Preliminary analysis of the role and significance of miR-9 in uveal melanoma invasiveness. MTT assay showed that genistein concentration-and time-dependent inhibition of growth of uveal melanoma cells. The treatment of 72-hour half of inhibition rate 48.23μM, and only 16.7% and high concentration 200μM cell viability. In vivo studies also showed that genistein treatment group can significantly inhibit the growth of xenografts in nude mice (F = 8.849; P = 0.001). Stem-loop real-time PCR results showed that genistein in a concentration-dependent inhibition of the expression of miR-27a. Compared with the control group, C918 cells after 25, 50, 100, and 200μM of genistein treatment, the expression levels of miR-27a were down to 88.9% ± 6.9%, 72.9% ± 5.4%, 51.7% ± 3.2% 42% ± 2.5%. Confirmed reduced miR-27a could ultimately affect the transcriptional activity of target genes, we detected simultaneously ZBTB10 expression, found that compared with the control group, ZBTB10 expression levels in genistein treatment group (200μM) was significantly higher (P = 0.037) . At the same time, we observed that compared with the highly invasive uveal melanoma C918 cells, the expression of miR-9 in minimally invasive uveal melanoma OCM-1A cells were significantly increased (P lt; 0.001), but also found that genistein C918 cell lines can be induced miR-9 expression (P = 0.022) implies the re-activation of the miR-9 may affect uveal melanoma invasion and metastasis. Therefore, in the present study, we first find the correlation genistein anticancer activity miRs regulation mechanism. The second part of the Pre-miR-27a gene polymorphisms and susceptibility to gastric cancer and mechanisms, miRNAs abnormal expression of many human cancers. highly conserved miRNA sequence, miRNA functional variation may affect a variety of biological processes. Therefore, when mutations or single nucleotide polymorphisms in miRNA genes affect the transcription of pri-miRNAs miRNAs mature miRNA-mediated transcriptional regulation. However miRNA gene polymorphism may affect the gastric susceptibility still unknown. In this study, we first compared miR-27a and its organization in gastric carcinoma and paraneoplastic target genes ZBTB10 expression differences and investigation occurred in the pre-miR-27a A / G (rs895819) polymorphism in the development of gastric cancer the role and significance, while observing the A / G variation affect the expression of miR-27a and target gene ZBTB10. Real-time quantitative PCR results showed that, compared with the adjacent normal tissue, miR-27a over-expression in gastric cancer tissues. The relative Ct rate of 78.79% ± 5.90% and 73.42 ± 4.80% (P = 0.023). In contrast to the (high low Ct-rate representatives of expression), compared with the adjacent normal tissue, ZBTB10 mRNA expression in gastric cancer tissues reduced. Was 100.30% ± 5.78% and 110.25% ± 5.98% (P lt; 0.001) relative Ct. Sources in the hospital, including 304 gastric cancer patients and 304 age and sex-matched control group, the study found that the G allele frequency in the case group was 39.97%, 32.73% was significantly higher (χ 2 = 6.88; P = 0.009). SNP rs895819 was significantly different from the distribution of genes in the case group and the control group (χ2 = 6.55; P = 0.038). The case group and the control group, the genotype distributions were in Hardy-Weinberg equilibrium (P = 0.194; P = 0.053). Compared with the AA gene carriers, the mutated gene (AG GG) subjects with high gastric cancer risk (adjusted OR, 1.48; 95% CI, 1.06-2.05; P = 0.019). Stratified analysis showed that this high-risk elderly subjects (gt; 58-year-old), male, non-smokers, patients of rural residents is particularly evident, also found that pre-miR-27a gene variation is associated with lymph node metastasis. In addition, in order to observe the pre-miR-27a mutant would affect the expression of miR-27a, we detected 65 cases of patients with gastric cancer tumor tissue expression of miR-27a, found that compared with the AA gene carriers AG GG gene carrying by miR-27a expression was significantly different from the rate was 78.98% ± 7.31% and 73.74% ± 4.34% (P lt; 0.001) relative Ct. However, the AG GG gene carriers ZBTB10 expression was significantly lower than the AA gene carriers. Compared with AA carriers, AG GG subjects had a higher rate of Ct (107.56% ± 7.05% and 101.68% ± 7.16%; P = 0.003). Our results indicate that pre-miR-27a, a common polymorphism (rs895819) through regulation miR-27a and ZBTB10 expression levels play an important role in gastric cancer susceptibility.

The Role of MicroRNAs and Target Gene in Uveal Melanoma and Gastric Cancer Development and Progression

Abstract

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