| Objective: Kruppel sample factor 4（Kruppel-like factor 4, KLF4） with a wide variety of tumor is closely related to the occurrence and development, in a different environment for the tumor has a dual regμlating function, KLF4 gene can inhibit the transcription process and can activate transcription occurs, both in the tumor can be used as a factor and may be a tumor suppressor factor to promote cancer. Accordingly, KLF4 expression exists in different tumors in the diversity, or play a role of oncogenes, or tumor suppressor genes. In this study we proposed test KLF4 MKN-45 in human gastric cancer cell line and normal gastric mucosa epithelial cell line GES-1 the expression, and combined with the bioinformatics analysis of KLF4 gene promoter area, predict possible transcription regμlatory factors in the process of gastric cancer occur, for further research on the transcriptional regμlatory mechanism-provides research foundation. Methods: Gastric cancer cell lines MKN-45 and normal gastric mucosa epithelial cell line GES-1, extraction cell lines of RNA, reverse transcription it into c DNA,using real-time fluorescent quantitative PCR method of the above two kinds of cells was detected in KLF4 m RNA expression level; Using bioinformatics method to obtain basic information KLF4 gene, the promoter sequence ID, to overcome the limitations of a single software, through Promotor Scan, Patch, P-Match, Ali Baba2 online prediction software analysis of KLF4 gene promoter sequences, to predict people KLF4 promoter region cis role element and related transcription factors, using gene expression profile database for protein abnormal expression in gastric cancer tissue, combining forecasting resμlts out transcription factor of KLF4 gene may be involved in gastric cancer at last. Resμlts: The real-time fluorescent quantitative PCR detection resμlts showed that KLF4 m RNA expression in human gastric cancer cell lines MKN- 45 below the expression in normal gastric mucosa cell lines GES-1（4.24±0.15 vs. 6.66±0.23）, statistically significant difference（p < 0.01）. Through bioinformatics analysis of KLF4 gene promoter regions, get people KLF4 gene ID: 9314, located in nine q31, total length of 4915 bp, the m RNA ID: NM004235. 4, the gene promoter ID: HPRM23684, total length of 1319 bp. These softwares analysis of KLF4 promoter sequence, in which Promotor Scan for 57 transcription factor binding sites, 17 transcription factors involved, including ATF, EGR-1, GCF, H4TF1, MLTF, MT-I.6, Sp1 etc.. Through Patch get 562 binding sites of transcription factor, involving 113 transcription factors,such as AP-1, H4TF1, LEF-1, NF-1, Pax-5, SRF, SMAD-4 etc.. The P-Match parameter is set to "to minimize the sum of both error rates", for 18 transcription factor binding sites, 9 transcription factors, including AP-4, ARP-1, ATF, NF-E2 and so on; parameter is set to "to minimize false negative matches", get 295 transcription factor binding sites, 25 transcription factors, including AP-4, ATF, CREB, NF-1, NF-E2, YY1 etc.. Ali Baba2 software for 172 binding sites of transcription factor, 52 transcription factors, including AP-2, ATF, NF-1, Sp1, USF, YY1 etc.. Summing up the above results, involved 183 transcription factors. In addition, through online gene expression profile database, abnormal expression of protein in gastric cancer, a total of 3249, compared with the 183 transcription factor for screening, prompt 17 transcription factors in human gastric cancer which may be involved in the regulation of KLF4, including LEF-1、NF-1、Pax-5、SMAD-4、SRF、YY1, etc. Conclusion: This study prompt detection in a hypoechoic KLF4 expression in gastric cancer cells MKN-45, bioinformatics analysis showed that the expression may be AP-1、AP-2、ATF、FXR、HIF-1、LEF-1、NF-1、Pax-5、RREB-1、SMAD-4、SP3、SRF、USF1 and YY1 transcription factor regμlation, including the transcription factors to promote the development of cancer with tumor suppression effect is also including some transcription factors. |
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