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Single-nucleotide Polymorphisms in MicroRNA Binding-sites Are Potentially Associated with the Cancer Risk and Prognosis for Hepatocellular Carcinoma

Author: WuChenSi
Tutor: ZhangRuiXing;GuoZhanJun
School: Hebei Medical University
Course: Internal Medicine
Keywords: Primary liver cancer MicroRNA Combined with Bit point Single nucleotide polymorphisms Survival rate
CLC: R735.7
Type: Master's thesis
Year: 2010
Downloads: 265
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Abstract


The purpose of: MicroRNAs (miRNA) are a class of small-scale non-coding RNA, is composed of a about of 22 nucleotide, its encoded by chromosomal DNA, at the posttranscriptional level to play a regulation and control the the the function of of the gene expression. A growing body and more research found that, miRNA has a the adjust the embryonic development, cell proliferation, differentiation, apoptosis and tumor occurrence of, and and so on the biological function of, miRNA mainly through the with the the the 3'UTR (3 of other target mRNA in 'in (on) a (non-) untranslated region) complete or incomplete the nucleotide of complementarily binding, in in order to to to to achieve its negative-sexual adjuster the features on of the of the the target mRNA, thereby regulating the the the Expression of of the gene becomes. Of miRNA in the the gene on the binding sites single nucleotide polymorphism (SNP) with the human tumors of occurrence of the event are closely related, but About the the research of the miRNA-binding-bit point SNP with the primary liver cancer the relationship between very little. In this study, intends to to through the on the the closely related to the with the tumor miRNA-binding-bit point SNP carried out sequencing analysis, and follow-up the in patients with the corresponding primary liver cancer, survival analysis was performed, Case report combined with - control study, identification out of the with the occurrence of hepatocellular carcinoma and prognosis of-related specificity SNP-bit point, in on the the analysis of the to to explore the the the the individual and the the the relationship between of the tumor susceptibility factors, screening the the of the high-risk groups of the susceptible to liver cancer, as well as to carried out a the prognosis of evaluation, the establishment of a effective early-warning mechanism great significance to both. Methods: 1 object of study: liver cancer group of patients for the September 2007 ~ inter-in March 2008, the the patients with of in the the the Fourth Hospital of of the of Hebei Medical University, liver and gallbladder surgical inpatients of the surgery, by the the and other tests of laboratory and imaging, clinical diagnosis as primary liver cancer, the line of primary liver cancer radical surgery, All patients with have their own confirm the diagnosis by postoperative and pathology to as a hepatocellular carcinoma, liver cancer the group of 60 cases of primary liver cells Any specimen of the of liver cancer of, are taken from the surgical resection of the tumor tissue; the 60 cases of of the control group anticoagulant venous blood specimens, all from the the in October 2008 ~ inter-in April 2009, the in the the the Fourth Hospital of of the of Hebei Medical University, acquisition of the normal subjects venous blood samples were. DNA of the 2 The Gene group extraction: use the the the Gene-group DNA of the Extraction of The the reagent cartridge was purified from DNA. Organizations and blood specimens of, After the extraction Save standby in the placed in the 4 ℃ In Fridge - thermostat Fridge. 3 PCR (polymerase chain REACTION) amplification of miRNA-binding-bit Points Point of Destination fragment: in the the Find a in the the BLAST server of the National Center for Biotechnology Information (NCBI) the the The purpose of gene of the with the 12 miRNA combination of corresponding to the SNP-bit point, to design PCR amplification primers, with a the PCR premixed kit (colorless system) were amplified by, PCR product was purified by the after the confirmed of the agarose gel electrophoresis, sent to the the sequencing of the Shanghai bio-engineering techniques Service Co., Ltd.. 4 follow-up: through the telephone to follow-up of the the survival of the situation of the postoperative of the in patients with primary liver cancer. 5 statistical analysis: the use of t measured in terms of test was used to compare the in information, using the of the liver cancer group with the the control group for the Electrochemically Modified chi-square inspection and analysis of the SNP distribution frequency of, the use of Kaplan-Meier method estimated to be the survival rate and draw a in the between the two groups is survival function curve, accumulated, with a of Log-Rank method on in each group the similarities and differences distribution of by the survival curves can be tested, comparison survival rate to have no difference in. All data analysis applications SPSS10.0 statistical software, P lt; 0.05 are thought to have statistically significant. Results: on the the combination of with the the of miRNA in the gene on the 12 SNP-bit Points Point of Destination fragment were amplified by and sequenced, found that the rs12583, rs1128665, rs17703261, rs14109, rs17107469, rs10463 six polymorphism-bit point, low-frequency allele the emergence of rate of is extremely small, Sequence analysis showed that the genotype single, not suitable for correlation analysis, screening out rs1044129a, rs4901706, rs16917496, rs1053667, rs11337, of rs3660 six polymorphisms point subsequent studies. 1 pairs of The combination of with the the of miRNA in the gene on the six the SNP distribution frequency of of the-bit point is analysis showed that In the between the the liver cancer group and control group, rs16917496, rs1053667, rs11337, inter-the in each group of the rs3660 polymorphism-bit points comparison, found that the This four -bit point SNP distribution frequency of was no significant difference. While the, For the RyR3 gene of polymorphic loci rs1044129a, A / A, the the A / G genotype frequencies of existence of difference (x2 = 5.562, P = 0.018), A / A, the A / GG / G genotype frequencies of there are differences (Chi-square = 4.261, P = 0.039); For the the of the of polymorphic loci rs4901706 of the C14orf101 gene, G / g, A / G genotype frequency there are differences (x2 = 5.022, P = 0.025), the, the A / GA of the G / G / A genotype frequency there are differences (chi2 = 5.647, P = 0.017), G, the A allele-type frequency there are differences (chi2 = 4.843, P = 0.028). Above results indicate, the the rs1044129a-bit of of the RyR3 gene the rs4901706-bit point of the the gene of point and the C14orf101 the SNP distribution frequency of there are differences, its related to occurrence of with the primary liver cancer. 2 pairs of combination of with the the of miRNA in the gene on the six SNP-bit point, the the the survival rate of of the in patients with primary liver cancer of the differences of all genotype for analysis, rs1044129a, rs4901706, rs16917496, rs11337, rs3660 five of polymorphic loci, The differences of all genotype patient group survival rates between the no significant difference. As for the the the of polymorphic loci of the KIAA0423 gene rs1053667, 2-year survival of the the genotype the patient's of the T / T (38), C / T (15), C / C (3), respectively, for the 44.74%, 40.00%, 0.00%, respectively, which the the between the two groups survival rate of the T / T, C / of patients with C genotype has there is a difference to (P = 0.0054). Combined with the survival function curve analysis, and the the other genotype of comparison, the C / C genotype the survival shortest period of of patients with, C / C genotype and survival rate of patients with was a negative correlation, C allele may be for the control of genetic, affect the in patients with primary liver cancer The survival rate of recessive genes. These results suggest that the KIAA0423 gene rs1053667 locus SNP and primary liver cancer prognosis. Conclusion: between liver cancer and control groups the RyR3 gene of polymorphic loci rs1044129a of the SNP and C14orf101 gene polymorphic loci rs4901706 SNP frequency distribution of differences, the two sites may be associated with primary liver cancer An the relevant the SNP-bit of point has occurred. 2 For the of polymorphic loci of the KIAA0423 gene rs1053667, T / T, C / C genotype patients with survival rate there is a difference, C / C genotype the survival shortest period of of patients with, and with the the the survival rate of patients was a negative correlation, this bit point C, etc. -bit gene may be for the control of of genetic, affect the the survival rate of the in patients with primary liver cancer was of the recessive gene, rs1053667-bit point may be as affecting the the point of the the the relevant SNP-bit of the primary liver cancer the prognosis of.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Liver tumors
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