TNRC9 基因rs12443621A/G 多态性与中国女性乳腺癌易感性及临床病 理关系的研究

目的:研究TNRC9/LOC643714 基因 rs12443621A/G 多态性与乳腺癌易感性及临床病理之间的关系.方法:DNA 试剂盒提取321 例乳腺癌患者和340 例正常女性静脉血全基因组DNA,PCR扩增目的基因片段,提取扩增样本进行DNA 测序检测分析rs12443621 多态性.应用 SPSS17.0 软件对实验结果进行统计学分析.结果:应用 SPSS17.0软件对TNRC9/LOC643714 基因rs12443621A/G多态性AA、AG、GG 进行卡方检验分析,结果显示三种基因型分布在病例组及对照组中无统计学意义(X<'2>=1.43,P>0.05),与乳腺癌易感性无关,与乳腺癌病理分型、ER、PR、HER-2状态以及淋巴结是否转移无相关性(X<'2>=2.90,P>0.05;X<'2>=2.25,P>0.05;X<'2>=1.671,P>0.05;X<'2>=1.34,P>0.05;X<'2>=3.24,P>0.05).结论:TNRC9 基因rs12443621A/G 多态性与乳腺癌易感性及临床病理特征无关.不能作为独立的基因标志物对乳腺癌进行早期检测和诊断.

Abstract：

Objective: To investigate the relationship between the rs12443621 polymorphisms of TNRC9/LOC643714 and breast cancer risk and clinico-pathological characteristics in Chinese women. Methods: Genomic DNA was extracted from peripheral blood. The Single Nucleotide Polymorphisms of the TNRC9/LOC643714 rs12443621,from breast cancer of 321 cases and 340 controls were detected by polymerase chain reaction (PCR) and direct DNA sequencing.Results:The genotypes of rs12443621 can not increase the risk for breast cancer(X2=1.43,P>0.05).There was no relationship between the genotypes and pathological category,lymph node metastases, ER status or PR staus or Her-2 status (x2=2.90,P>0.05; X2=2.25,P>0.05;X2=1.671,P>0.05; X2=1.34,P>0.05; X2=3.24,P>0.05). Conelusion:There was no relationship between three genotypes of rs12443621A/G and individual susceptibitity or clinic pathological characteristics of breast cancer.     

长链非编码RNA GAS5抑制内皮细胞功能

最新研究表明,长链非编码RNA GAS5（lncRNA GAS5）可调节血管内皮细胞的凋亡,但对内皮细胞其他功能的调控并不明确。本研究旨在了解lncRNA GAS5对内皮细胞的增殖、成血管、NO分泌及内皮标志分子CD31和vWF表达的影响及可能机制。将LncRNA GAS5干扰慢病毒（LV-GAS5-RNAi）转染人脐静脉内皮细胞株（EA.hy926）后,采用CCK8及Matrigel胶分别检测EA.hy926的增殖和成血管能力;硝酸还原酶法检测NO的分泌情况;real-time RT-PCR检测CD31、vWF及miR-21的表达;Western印迹检测PTEN在蛋白质水平的表达。结果显示：与对照组比较,LV-GAS5-RNAi组EA.hy926增殖能力无明显变化(0.34±0.01 vs. 0.34±0.04,P>0.05),而其成血管能力升高(133.70±12.64 vs. 100.00±4.65,P<0.05),NO的分泌量亦增加(28.54±2.75 μmol/L vs.15.11±1.19 μmol/L,P<0.01);内皮标志分子CD31（是对照组的1.46倍）及vWF（是对照组的2.94倍)的基因表达量均显著升高。同时,miR-21表达亦明显升高(是对照组的1.42倍),而miR-21下游靶基因PTEN蛋白质的表达量则显著降低（0.13±0.05 vs. 0.38±0.03,P<0.01）。以上结果提示,LncRNA GAS5抑制了内皮细胞的功能,miR-21、PTEN信号分子可能参与其中的调节。     

Exploring a possible association between the occurrence of the SERPINE1-675 4G/5G (rs1799889) polymorphism and the increased risk of esophageal cancer in the Caucasian population

The goal of this research was to analyze the SERPINE1 -675 4G/5G (rs1799889) and MMP9 T-1702A (rs2297864) polymorphisms in esophageal cancer among polish patients, classified as part of the Caucasian population. The analysis of polymorphic gene variants was performed on 35 randomly selected samples excised from patients with esophageal cancer. The tissue specimens were stored as Formalin-Fixed, Paraffin-Embedded (FFPE) blocks. All patients in the sample group were of Caucasian ethnicity. The genotype distribution of MMP9 T-1702A and SERPINE1 -675 polymorphisms was analyzed using the Restriction Fragment Length Polymorphism (RFLP) method. A correlation between the expression of ?675 polymorphic form of SERPINE1 and alcohol abuse has been found. Additionally, a correlation between the ?675 polymorphism and the subtype of EC developed by the patient has been shown. To the best of the authors’ knowledge, this is the first report investigating the SERPINE1 -675 4G/5G (rs1799889) polymorphism as a potential candidate for a prognostic biomarker of esophageal cancer.     

CHRNA5 variants moderate the effect of nicotine deprivation on a neural index of cognitive control

Individuals with reduced attention and memory cognitive control‐related processes may be motivated to smoke as a result of the cognitive enhancing effects of nicotine. Further, nicotine deprivation‐induced reductions in cognitive control may negatively reinforce smoking. Minor allele carriers at rs16969968 in the nicotinic acetylcholine receptor α5 subunit gene (CHRNA5) have been shown to exhibit both reduced cognitive control and greater nicotine dependence. It is therefore of interest to see if variants in this gene moderate the influence of nicotine deprivation on cognitive control. P3b and P3a components of the event‐related brain potential waveform evoked by a three‐stimulus visual oddball task are widely viewed as positive indices of cognitive control‐related processes. We tested the hypothesis that individuals possessing at least one minor allele at rs16969968 in CHRNA5 would show greater nicotine deprivation‐induced reductions in P3b and P3a amplitude. The sample included 72 non‐Hispanic, Caucasian heavy smokers (54 men and 18 women) with a mean age of 36.11 years (SD = 11.57). Participants completed the visual oddball task during counterbalanced nicotine and placebo smoking sessions. Findings indicated that rs16969968 status did not moderate nicotine effects on P3b or P3a, whereas variation in other CHRNA5 polymorphisms, which are not as well characterized and are not in linkage disequilibrium with rs16969968, predicted nicotine deprivation‐induced reduction of P3a amplitude: rs588765 (F_1,68 = 7.74, P = 0.007) and rs17408276 (F_1,67 = 7.34, P = 0.009). Findings are interpreted in the context of vulnerability alleles that may predict nicotine effects on cognitive control.     

IL28B gene polymorphisms and Th1/Th2 cytokine levels might be associated with HTLV-associated arthropathy

The present study is the first investigation of the association between single nucleotide polymorphisms (SNPs – rs8099917, rs12979860 and rs8103142) of the IL28B gene and the development of human T-lymphotropic virus (HTLV)-associated arthropathy (HAA). Individuals with HAA exhibited low interleukin (IL) 6 (p < 0.05) and high IL-10 (p < 0.05) levels compared with asymptomatic patients. TNF-α/CD4⁺ T cell count, TNF-α/CD8⁺ T cell count and IFN-γ/proviral load positively correlated in asymptomatic patients. The allelic and genotypic frequencies did not differ between patients with HAA and asymptomatic patients. Seven haplotypes were detected in the investigated population, with haplotype CCT (p < 0.05) being the most frequent among the HTLV-infected individuals, while haplotype TTG (p < 0.05) was detected in the group with HAA only. Compared with asymptomatic patients, individuals with HAA and genotype TT (rs8099917) exhibited larger numbers of CD8⁺ T cells (p < 0.05) and higher proviral load levels (p < 0.05). Those patients with HAA and genotypes CC (rs12979860) and TT (rs8103142) exhibited high TNF-β (p < 0.05) and IFN-γ (p < 0.05) levels. Those patients with HAA and genotype CT/TT (rs12979860) exhibited high IL-10 levels (p < 0.05). These results suggest that haplotypes CCT and TTG might be associated with susceptibility to HTLV infection and progression to HAA, respectively. Genotype TT (rs8099917) might be a risk factor for elevation of the proviral load and CD8⁺ T cell count. In addition, genotypes CC (rs12979860) and TT (rs8103142) seem to be associated with increased TNF-β and IFN-γ levels.     

Hypoxia stabilizes GAS6/Axl signaling in metastatic prostate cancer

The receptor tyrosine kinase Axl is overexpressed in a variety of cancers and is known to play a role in proliferation and invasion. Previous data from our laboratory indicate that Axl and its ligand growth arrest-specific 6 (GAS6) may play a role in establishing metastatic dormancy in the bone marrow microenvironment. In the current study, we found that Axl is highly expressed in metastatic prostate cancer cell lines PC3 and DU145 and has negligible levels of expression in a nonmetastatic cancer cell line LNCaP. Knockdown of Axl in PC3 and DU145 cells resulted in decreased expression of several mesenchymal markers including Snail, Slug, and N-cadherin, and enhanced expression of the epithelial marker E-cadherin, suggesting that Axl is involved in the epithelial-mesenchymal transition in prostate cancer cells. The Axl-knockdown PC3 and DU145 cells also displayed decreased in vitro migration and invasion. Interestingly, when PC3 and DU145 cells were treated with GAS6, Axl protein levels were downregulated. Moreover, CoCl(2), a hypoxia mimicking agent, prevented GAS6-mediated downregulation of Axl in these cell lines. Immunochemical staining of human prostate cancer tissue microarrays showed that Axl, GAS6, and hypoxia-inducible factor-1α (Hif-1α; indicator of hypoxia) were all coexpressed in prostate cancer and in bone metastases compared with normal tissues. Together, our studies indicate that Axl plays a crucial role in prostate cancer metastasis and that GAS6 regulates the expression of Axl. Importantly, in a hypoxic tumor microenvironment Axl expression is maintained leading to enhanced signaling.     

Association of caspase 8 promoter variants and haplotypes with the risk of breast cancer and its molecular profile in an Iranian population: A case-control study

Caspase 8 (CASP8) gene plays a key role in the regulation of apoptotic cell death. Expression variation in this gene has been associated with the risk of breast cancer. The aim of this study was to investigate the association of rs3834129 and rs3769821, as functional variants, and their haplotypes with molecular profile as well as the risk of breast cancer in an Iranian population. A case-control study was conducted on 812 participants including 293 breast cancer patients and 519 healthy controls. Genotyping was performed by polymerase chain reaction–based methods. Statistical analysis was performed using SPSS Ver16. The association between polymorphisms and haplotypes with the risk of breast cancer was estimated by calculating odds ratios (OR) and chi-square (χ²) tests. In comparison with ins allele (I) of rs3834129, carriers of del allele (D) showed a lower risk of breast cancer (OR, 0.65; 95% confidence interval [CI], 0.49-0.87; P = 0.004). The multivariate logistic regression model indicated DD genotype as an independent factor for a decreased risk of breast cancer in our population (OR, 0.18; 95% CI, 0.06-0.58; P = 0.004). Also, the C allele of rs3769821 was associated with a 43% increased risk of breast cancer (P = 0.005); however, after adjustment for confounding factors, no association with rs3769821 and breast cancer was observed. In addition, D-T haplotype and diplotype presented protective effects (P < 0.05). Our results indicate that genetic variations in the promoter region of CASP8 gene, especially rs3834129, may serve as a genetic risk factor for breast cancer in an Iranian population.     

COVID-19 Survivor Patients Carrying the Rs35705950 Risk Allele in MUC5B Have Higher Plasma Levels of Mucin 5B

Background: Genetic susceptibility to infectious diseases is partly due to the variation in the human genome, and COVID-19 is not the exception. This study aimed to identify whether risk alleles of known genes linked with emphysema (SERPINA1) and pulmonary fibrosis (MUC5B) are associated with severe COVID-19, and whether plasma mucin 5B differs according to patients’ outcomes. Materials and methods: We included 1258 Mexican subjects diagnosed with COVID-19. We genotyped rs2892474 and rs17580 of the SERPINA1 gene and rs35705950 of MUC5B. Based on the rs35705950 genotypes, mucin 5B plasma protein levels were quantified. Results: Homozygous for the risk alleles of the three polymorphisms were found in less than 5% of the study population, but no statistically significant difference in the genotype or allele association analysis. At the protein level, non-survivors carrying one or two copies of the risk allele rs35705950 in MUC5B (GT + TT) had lower levels of mucin 5B compared to the survivors (0.0 vs. 0.17 ng/mL, p = 0.0013). Conclusion: The polymorphisms rs28929474 and rs17580 of SERPINA1 and rs35705950 of MUC5B are not associated with the risk of severe COVID-19 in the Mexican population. COVID-19 survivor patients bearing one or two copies of the rs35705950 risk allele have higher plasma levels of mucin 5B.     

Association of genetic variants rs641153 (CFB), rs2230199 (C3), and rs1410996 (CFH) with age-related macular degeneration in a Brazilian population

This study aimed to investigate the association among genetic variants of the complement pathway CFB R32Q (rs641153), C3 R102G (rs2230199), and CFH (rs1410996) with age-related macular degeneration (AMD) in a sample of the Brazilian population. In a case-control study, 484 AMD patients were classified according to the clinical age-related maculopathy grading system (CARMS) and compared to 479 unrelated controls. The genetic variants rs1410996 of complement H (CFH), rs641153 of complement factor B (CFB), and rs2230199 of complement 3 (C3) were evaluated through polymerase chain reaction (PCR) and direct sequencing. The associations between single nucleotide polymorphisms (SNPs) and AMD, adjusted by age, were assessed by using logistic regression models. A statistically significant association was observed between AMD risk and rs2230199 variant with an OR of 2.01 (P  = 0.0002) for CG individuals compared to CC individuals. Regarding the comparison of advanced AMD versus the control group, the OR was 2.12 (P = 0.0036) for GG versus AA genotypes for rs1410996 variant. Similarly, the OR for rs2230199 polymorphism was 2.3034 (P  = 5.47^e-05) when comparing CG individuals to CC carriers. In contrast, the rs641153 variant showed a significant protective effect against advanced AMD for GA versus GG genotype (OR = 0.4406; P  = 0.0019). When comparing wet AMD versus controls, a significant association was detected for rs1410996 variant (OR = 2.16; P  = 0.0039) comparing carriers of the homozygous GG versus AA genotype, as well as in the comparisons of GG (OR = 3.0713; P  = 0.0046) and CG genotypes (OR = 2.2249; P  = 0.0002) versus CC genotype for rs2230199 variant, respectively. The rs641153 variant granted a significant protective effect against wet AMD for GA versus GG genotypes (OR = 0.4601; P  = 0.0044). Our study confirmed the risk association between rs2230199 and rs1410996 variants and AMD, and the protective role against AMD for rs641153 variant.     

Association of RNASEL and 8q24 variants with the presence and aggressiveness of hereditary and sporadic prostate cancer in a hispanic population

To study the association between the polymorphisms Arg462Gln and Asp541Glu from the RNASEL gene (1q25), and the polymorphisms rs620861, rs1447295, rs6983267, rs7837328 from the chromosome 8q24 with the risk of presenting prostate cancer (PCa) and its clinical characteristics in a Hispanic (Chilean) population. The study was performed on 21 control patients and 83 patients diagnosed with PCa. Polymorphisms were analysed from blood samples through real‐time PCR by using TaqMan probes, and the genetic analysis was performed with the SNPStats program. Also, a comparison was performed between clinical characteristics of PCa and the presence of the different polymorphism genotypes by using the Minitab software. There was a significant association between the genotype G/G from the polymorphism rs6983267 with an overall increased risk of PCa, in patients both with or without family history of PCa (OR = 4.47, 95% CI = 1.05–18.94, P = 0.034 and OR = 3.57, 95% CI = 0.96–13.35, P = 0.037, respectively). Regarding clinical parameters, patients carrying the genotype C/C from the polymorphism Asp541Glu had significantly higher prostate‐specific antigen (PSA) levels than patients carrying the other genotypes (P = 0.034). Moreover, patients with the genotype G/G of rs6983267 had higher PSA levels (P = 0.024). The polymorphism rs6983267 from region 3 of the chromosome 8q24 appears to be a prominent risk factor for PCa and a biomarker for cancer aggressiveness in the group of patients who presented higher levels of PSA at the time of diagnosis.     

Association of chemotactic chemokine ligand 5 rs2107538 polymorphism with tuberculosis susceptibility: A meta-analysis

A meta-analysis was carried out in this study by summarizing relevant research to evaluate the relationship between rs2107538 polymorphism in the chemotactic chemokine ligand 5 (CCL5) gene and tuberculosis (TB) susceptibility. Published studies were retrieved from PubMed, Embase, and CNKI databases using the keywords ‘CCL5’, ‘TB’, and ‘polymorphism’. Nine studies involving 2584 patients with TB and 2265 controls were included in the current meta-analysis. The combined results suggested that the CCL5 rs2107538 polymorphism was correlated with TB susceptibility (recessive model: OR = 1.45, 95% CI = 1.02–2.07). Subgroup analysis according to race indicated that such correlation could be detected in Caucasians (CT versus CC: OR = 1.53, 95% CI = 1.20–1.95; dominant model: OR = 1.58, 95% CI = 1.25–1.99), but not in East Asian, South Asian, and South African populations. In conclusion, the results of our meta-analysis suggest that CCL5 rs2107538 polymorphism might contribute to the risk of TB, especially in Caucasians. Well-designed studies with more subjects will be required for further validation of these results.     

Association study of rs10768683 and rs968857 polymorphisms with transfusion-dependent thalassemia (TDT) in a southern Iranian population

Previous studies reported that detection of polymorphisms inherited through paternal model could be potential markers for the Non-Invasive Prenatal Diagnosis (NIPD) of β-thalassemia. The aim of the current study was to find out the associations of rs10768683 and rs968857 with transfusion-dependent thalassemia (TDT) in a southern Iranian population. A total of 175 subjects were investigated, divided into patients with TDT as case group (n?=?75) and healthy people as control group (n?=?100). Genomic DNAs were extracted from peripheral blood using salting out procedure. Genotyping rs10768683 and rs968857 was carried out by ARMS-PCR, then statistical analyses were assessed using SPSS, and Medcalc ver. 18 software. Data showed that rs10768683 was statistically significant in co-dominant model of inheritance (P?=?0.025, OR?=?2.11 [1.08-4.15]) and genotype frequencies of CG among controls and cases were 0.68 and 0.80, respectively. However, according to genotype frequencies, there was no association between rs968857 and TDT among cases and healthy controls in any models of inheritance. In conclusion, the present study showed the association of rs10768683 with major β-thalassemia through ARMS-PCR technique.     

炎性肠病易感基因GPR35在肠炎发生发展中的功能研究

炎性肠病在全球范围内发生极其普遍,具有反复发作、难以治愈的特点,也是诱发结直肠癌的高风险因素之一。肠炎的发生与遗传因素密切相关,有报道发现位于GPR35基因座上的多个单核苷酸多态性(single nucleotidepolymorphism,SNP)位点rs4676410、rs3749171和rs3749172与肠炎敏感性高度相关,但是GPR35基因在肠炎的发生发展进程中的功能及相关机制尚没有明确结论。为了研究GPR35在肠炎中的作用,首先通过CRISPR/Cas9技术构建Gpr35敲除小鼠,随后利用DSS诱导的肠炎模型评价Gpr35在肠炎发生中的作用,发现敲除小鼠在体重变化、DAI评分、肠上皮损伤以及炎性细胞浸润等肠炎相关指标显著低于野生型小鼠。为了研究肠炎相关SNP突变对GPR35活性的影响,首先根据rs3749171和rs3749172SNP位点突变信息构建GPR35-T108M和GPR35-S294R两种突变型受体,其次通过多种GPR35下游信号通路活性测试,发现两种突变均能够增强GPR35受体活性。最后通过Westernblotting分析发现相较于野生型小鼠,Gpr35敲除小鼠肠上皮Erk1/2磷酸化水平增加,表明Gpr35敲除后可能通过上调Erk1/2信号通路的方式抑制肠炎的发生发展。综上所述,本研究发现人类肠炎易感的rs3749171和rs3749172位点可能通过激活GPR35及下游信号通路的方式促进肠炎的发生发展,为炎性肠病的治疗提供了潜在的药物作用靶点。     

5 Alpha-reductase in the non-secretory cells of the rat ventral prostate epithelium

Kinetic constants for the 5 alpha-reductase were determined in freshly isolated epithelial cells from the rat ventral prostate. Studies were also performed on stromal tissue but not isolated stromal cells for comparison. Secretory and non-secretory epithelial cells were separated by centrifugation in a Percoll gradient. Both epithelial cell populations metabolized testosterone to predominantly 5 alpha-dihydrotestosterone (5 alpha-DHT), although when expressed per cell the capacity for conversion was 3-4-fold higher for secretory cells (7.4 pmol/min/10(6) cells) than for non-secretory cells (2.3 pmol/min/10(6) cells; P less than 0.01 in 4 separate studies). When compared per mg cytosol protein this difference became non-significant. Stromal tissue contained a 5 alpha-reductase Vmax (expressed) per mg protein) which was comparable to the non-secretory cell enzyme. Lineweaver-Burke plots revealed different Km values for the different cell populations (12.5, 5.9 and 4.7 microM for secretory, non-secretory and stromal cells, respectively) suggesting the presence of different isoforms of the enzyme, or differences in the intracellular concentrations of enzyme antagonists.     

Upregulation of lncRNA GAS5 inhibits the growth and metastasis of cervical cancer cells

This study aims to figure out the methylation of long non-coding RNA GAS5 promoter in cervical cancer and the mechanism of GAS5 on the progression of cervical cancer cells. The expression of GAS5 and methylation state of GAS5 in cervical cancer tissues and cells were determined. With the aim to to explore the ability of GAS5 in the proliferation, cell cycle progression, apoptosis, invasion, migration as well as the tumor growth, and metastasis in nude mice were determined. The expression of GAS5 was decreased and methylation state of GAS5 was elevated in cervical cancer. Overexpression of GAS5 inhibited proliferation, cell cycle progression, invasion, migration while inducing apoptosis of cervical cancer cells as well as suppressed tumor growth and metastasis in nude mice. Our study demonstrates that abnormal methylation of GAS5 contributes to poor expression of GAS5 in cervical cancer. In addition, upregulation of GAS5 inhibits the cervical cancer development.