• Asian Pacific Journal of Cancer Prevention
• /
• 제15권3호
• /
• pp.1181-1186
• /
• 2014

Sialyltransferase gene expression is altered in several cancers, including examples in the cervix. Transcriptional regulation of the responsible genes depends on different promoters. We aimed to determine the association of single-nucleotide polymorphisms in the B3 promoter of the ST3GAL4 gene and the P1 promoter of the ST6GAL1 gene with cervical premalignant lesions or cervical cancer. A blood sample and/or cervical scrapes were obtained from 104 women with normal cytology, 154 with premalignant lesions and 100 with cervical cancer. We also included 119 blood samples of random donors. The polymorphisms were identified by sequencing from PCR products. For the B3 promoter, a fragment of 506 bp (from nucleotide -408 to +98) was analyzed, and for the P1 promoter a 490 bp (-326 to +164) fragment. The polymorphism analysis showed that at SNP rs10893506, genotypes CC and CT of the ST3GAL4 B3 promoter were associated with the presence of premalignant lesions (OR=2.89; 95%CI 1.72-4.85) and cervical cancer (OR=2.23; 95%CI 1.27-3.91). We detected only one allele of each polymorphism in the ST6GAL1 P1 promoter. We did not detect any genetic variability in the P1 promoter region in our study population. Our results suggest that the rs10893506 polymorphism -22C/T may increase susceptibility to premalignant and malignant lesions of the cervix.

• IMMUNE NETWORK
• /
• 제5권4호
• /
• pp.205-214
• /
• 2005

Background: We examined global gene expression profiles of peripheral blood mononuclear cells (PBMCs) in patients with ulcerative colitis (DC), and tested whether the identified genes with the altered expression might be associated with susceptibility to UC. Methods: PBMCs from 8 UC and 8 normal healthy (NH) volunteers were collected, and total RNAs were subjected to the human 8.0K cDNA chip for the micro array analysis. Real time-PCR (RT-PCR) was performed to verify the results of micro array. One hundred forty UC patients and 300 NH controls were recruited for single nucleotide polymorphism (SNP) analysis. Results: Twenty-five immune function-related genes with over 2-fold expression were identified. Of these genes, two chemokines, namely, CXCL1 and CCL20, were selected because of their potential importance in the evocation of host innate and adaptive immunity. Four SNPs were identified in the promoter and coding regions of CXCL1, while there was no significant difference between all patients with UC and controls in their polymorphisms, except minor association at g.57A>G (rs2071425, p=0.02). On the other hand, among three novel and one known SNPs identified in the promoter region of CCL20, g. -1,706 G>A (p=0.000000055), g. -1,458 G>A (p=0.0048), and g. -962C>A (p=0.0006) were found to be significantly associated with the susceptibility of Uc. Conclusion: Altered gene expression in mononuclear cells may contribute to IBD pathogenesis. Although the findings need to be confirmed in other populations with larger numbers of patients, the current results demonstrated that polymorphisms in the promoter region of CCL20 are positively associated with the development of Uc.

• 생명과학회지
• /
• 제17권8호통권88호
• /
• pp.1157-1165
• /
• 2007

포도 ASR (VvMSA) 단백질은 hexose transporter 유전자 VvHT1의 전사를 조절하는 조절 인자 중의 하나로서 sugar 및 abscisic acid (ABA) 신호에 의해 발현이 유도된다. 본 연구진은 ACP RT-PCR (annealing control primer reverse transcriptase-polymerase chain reaction) 방법을 이용하여 포도 과실발달 과정에서 조절되는 유전자 중 VvMSA와 동일한 cDNA (VlASR)를 클로닝하였다. 이 유전자는 착과 시기에 발현되기 시작하여 과실이 발달하면서 점점 증가하여 착과 후 10 주에 가장 많이 발현되며, 숙기 후반에는 도리어 발현양이 감소하였다. 포도 asr 유전자의 조절기작을 밝히기 위해, 이 유전자의 genomic clone을 분리하였다. 총 1375 bp로 이루어진 이 유전자 절편에는 open reading frame과 100 bp의 intron을 포함하고 있다. 약 600 bp 길이의 프로모터 내에는 sugar 신호전달과 연관이 있는 것으로 알려진 sugar box(sucrose box 3 +sucrose response box 1)가 있다. 프로모터 절편을 reporter 유전자와 연결하여 Arabidopsis에 도입하고 형질전환체를 분석한 결과, reporter 유전자는 sucrose 처리와 상관없이 항상 발현되었다. 이러한 결과는 포도에서 보고된 ASR/VvHT1를 매개로 하는 sugar/ABA 신호전달계가 asr 유전자가 없는 Arabidopsis에서는 작동되지 않음을 시사하고 있다.

• Journal of Animal Science and Technology
• /
• 제53권1호
• /
• pp.15-22
• /
• 2011

자돈의 생시 체중은 자돈의 이유 체중 및 생존율에도 크게 영향을 미친다. 또한 출하 시까지 돼지의 성장률 뿐만아니라 출하 일령 단축 및 출하 체중과도 밀접한 연관성이 있는 것으로 알려져 있다. 본 연구는 돼지 7번 염색체의 PGK 2 (phosphoglycerate kinase 2) 유전자의 promoter 영역 및 transcription 영역에 해당하는 DNA 염기서열을 유전체 구조분석을 통해 20개의 SNP (Single Nucleotide Polymorpism)를 발굴하였고, 발굴된 SNP 중 PGK 2 유전자의 전사 조절 영역 내 g.122 T>G 다형을 재래돼지와 Landrace를 이용한 $F_2$ 집단 268두 자돈의 성장 형질과의 연관성 분석을 실시한 결과, 생시 체중(p<0.01) 및 3주령 체중(p<0.001)에서 통계적으로 고도의 유의적 연관성이 있음을 확인 할 수 있었다. 따라서 본 연구를 통하여 확인된 PGK 2 유전자의 promoter 영역 내 성장 형질 관련 SNP는 건강한 자돈 및 종돈을 조기 선발하기 위한 유전자 marker로 활용 가능성을 제시하였다.

• Journal of Plant Biotechnology
• /
• 제32권3호
• /
• pp.209-215
• /
• 2005

락토페린은 철 결합 당단백질로서 항 미생물활성과 면역강화와 같은 생리활성 기능을 가지고 있다. 본 연구는 배양 세포 고 발현 SWPA2 promoter를 이용하여 인체락토페린(hLf)을 생산하는 형질전환 가시오갈피 배양세포주 개발에 관한 것이다. 형질전환에 이용된 벡터는 산화스트레스 유도성 SWPA2 promoter의 조절 하에서 hLf이 소포체로 targeting되도록 제작된 SWPA2pro::ER-hLf/pCAMBIA이다. hLf을 생산하는 각 형질전환 배양세포들은 PCR과 Southern분석을 통해 hLf 유전자가 가시오갈피 게놈내로 성공적으로 도입되었음을 확인하였으며, western blot과 ELISA를 통해 형질전환 가시오갈피 배양 세포주에서 hLf 단백질이 활성이 있음을 확인하였다. 형질전환 가시오갈피 배양세포에서 hLf 단백질의 함량은 세포배양이 진행될수록 증가하여 정지기 때 가장 높았으며 전체 수용성 단백질의 약 3%를 차지하였다. 따라서 본 연구에서 개발된 인체락토페린을 고생산하는 약용식물 가시오갈피 배양 세포주는 산업적으로 이용될 수 있을 것으로 기대된다.

• 식물조직배양학회지
• /
• 제25권3호
• /
• pp.147-152
• /
• 1998

AL1-gene은 TGMV의 복제에 매우 중요한 역할을 하고 있다. 이 AL1 gene의 발현을 억제하기 위해서는 식물체내에서 AL1 gene의 antisense RNA의 발현에 의한 억제가 효과적 방법 중에 하나로 알려져 있다. 이런 발현을 식물체내에서 실현시키기 위해 hygromycin 저항성 유전자에 antisense AL1-gene을 연결시키고, 연결된 부위를 CaMV35s-promoter와 octopine synthase gene terminator 사이에 연결시켰다. 이 유전자 발현 단위 부분을 다시 kanamycin 저항성 유전자 발현 단위 부분을 지니고 있는 형질 전환 벡터인 pBinAR에 삽입시켜 새로운 형질 전환 벡터인 pAR35-2를 개발하였다. 이 벡터를 Agrobacterium tumefaciens LBA4404에 형질전환 시킨 다음, 토마토와 담배 잎사귀 조직에 감염시켜 식물체들을 kanamycin과 hygromycin이 함유된 배지위에서 배양하여 형질전환된 식물체들을 선발하였다. 형질 전환된 식물체들로부터 antisense AL1-gene 및 antisense RNA를 각각 PCR 및 RT-PCR를 이용한 southern hybridization 방법을 이용하여 증명하였고, 토마토 식물체의 공변세포쌍 내에 있는 엽록체 숫자가 여덟 개라는 것이 확인되어 형질 전환된 토마토 식물체가 2 배수체로서 정상적인 식물체라는 것을 증명하였다. 이러한 형질 전환 식물체는 앞으로 항 바이러스성 형질을 지니는 식물체들을 개발하는 데 많은 도움을 주리라 여겨 진다. 그리고, 본 연구에서 제조된 벡터 pAR35-2는 두 개의 항생제에서 동시 선발 할 수 있도록 되어 있고 promoter가 두 개로 되어 있어 형질 전환 식물체선발 및 유전자 발현 연구에 효과적으로 이용되어 질 수 있으리 라 여겨진다.

• BMB Reports
• /
• 제40권5호
• /
• pp.625-635
• /
• 2007

The enzyme squalene synthase (EC 2.5.1.21) catalyzes a reductive dimerization of two farnesyl diphosphate (FPP) molecules into squalene, a key precursor for the sterol and triterpene biosynthesis. A full-length cDNA encoding squalene synthase (designated as TcSqS) was isolated from Taxus cuspidata, a kind of important medicinal plants producing potent anti-cancer drug, taxol. The full-length cDNA of TcSqS was 1765 bp and contained a 1230 bp open reading frame (ORF) encoding a polypeptide of 409 amino acids. Bioinformatic analysis revealed that the deduced TcSqS protein had high similarity with other plant squalene synthases and a predicted crystal structure similar to other class I isoprenoid biosynthetic enzymes. Southern blot analysis revealed that there was one copy of TcSqS gene in the genome of T. cuspidata. Semi-quantitative RT-PCR analysis and northern blotting analysis showed that TcSqS expressed constitutively in all tested tissues, with the highest expression in roots. The promoter region of TcSqS was also isolated by genomic walking and analysis showed that several cis-acting elements were present in the promoter region. The results of treatment experiments by different signaling components including methyl-jasmonate, salicylic acid and gibberellin revealed that the TcSqS expression level of treated cells had a prominent diversity to that of control, which was consistent with the prediction results of TcSqS promoter region in the PlantCARE database.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권9호
• /
• pp.3921-3925
• /
• 2014

RASSF1A has been reported to be a candidate tumor suppressor in esophageal squamous cell carcinoma (ESCC). However, the association between RASSF1A promoter methylation and ESCC remains unclear. Eligible studies were identified through searching PubMed, Medline, Web of Science, and the China National Knowledge Infrastucture database. Studies were pooled and odds ratios (ORs) with corresponding confidence intervals (CIs) were calculated. Funnel plots were also performed to evaluate publication bias. Twelve studies involving 859 cases and 675 controls were included in this meta-analysis. A significant association was observed between RASSF1A methylation and ESCC overall (OR = 11.7, 95% CI: 6.59-20.9, z=8.36, P<0.00001). Subgroup analysis showed that the OR for heterogeneous tissues was 5.35 (95% CI = 2.95-9.71) while for autologous tissues it was 16.0 (8.31-30.96). For patient sample size, the OR for the <50 subgroup was 9.92 (95% CI = 2.88-34.2) and for the 50 case group was 13.1 (95% CI = 6.59-25.91). The OR for a relationship between RASSF1A methylation and TNM stages was 0.27 (95% CI=0.10-0.77), whereas there were no significant differences in RASSF1A methylation in relation to gender and differentiation among ESCC cases. This meta-analysis suggests a significant association between RASSF1A methylation and ESCC.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권11호
• /
• pp.4563-4566
• /
• 2014

Background: To evaluate relationship between the cyclooxygenase-2 promoter 765G/C polymorphism and digestive cancer risk in China. Materials and Methods: A literature search through February 2014 was performed using PubMed, Chinese Biomedical Literature Database (CBM) and China National Knowledge Infrastructure (CNKI) databases, and a meta-analysis was performed with RevMan 5.2 software for odds ratios and 95%CIs. Results: In total, 9 articles with 3,263 cases and 4,858 controls were included in this meta-analysis. The pooled OR (95%CIs) in the co-dominant model (GC vs GG) was 1.56 [1.19, 2.06], and in the dominant model ((CC+GC) vs GG), the pooled OR was 1.59 [1.21, 2.09] in overall cancers. In the subgroup analysis, stratified by cancer type, significant associations were found that the-765C allele had increased pancreatic cancer and gastric risk. No significant liver cancer and colorectal cancer risk of COX-2 -765G/C polymorphism was found. Conclusions: These findings suggest that COX-2-765*C is related to cancer susceptibility and may increase gastric and pancreatic cancer risk.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권6호
• /
• pp.3719-3724
• /
• 2013

Background: RASSF1A has been reported to be a candidate tumor suppressor in non-small cell lung cancer (NSCLC). However, the association between RASSF1A promoter methylation and NSCLC remains unclear, particularly in regarding links to clinicopathologic features. Methods: Eligible studies were identified through searching PubMed, EMBASE, Cochrane Library and China National Knowledge Infrastructure (CNKI) databases. Studies were pooled and odds ratios (ORs) with corresponding confidence intervals (CIs) were calculated. Funnel plots were also performed to evaluate publication bias. Results: Nineteen studies involving 2,063 cases of NSCLC and 1,184 controls were included in this meta-analysis. A significant association was observed between RASSF1A methylation and NSCLC in the complete data set (OR = 19.42, 95% CI: 14.04-26.85, P < 0.001). Pooling the control tissue subgroups (heterogeneous/autologous) gave pooled ORs of 32.4 (95% CI, 12.4-84.5) and 17.7 (95% CI, 12.5-25.0) respectively. Racial subgroup (Caucasian/Asian) analysis gave pooled ORs of 26.6 (95% CI, 10.9-64.9) and 20.9 (95% CI, 14.4-30.4) respectively. The OR for RASSF1A methylation in poorly-differentiated vs. moderately/well-differentiated NSCLC tissues was 1.88 (95% CI, 1.32-2.68, P<0.001), whereas there were no significant differences in RASSF1A methylation in relation to gender, pathology, TNM stage and smoking behavior among NSCLC cases. Conclusion: This meta-analysis suggests a significant association between RASSF1A methylation and NSCLC, confirming the role of RASSF1A as a tumor suppressor gene. Large-scale and well-designed case-control studies are needed to validate the associations identified in the present meta-analysis.