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| Methodology Research on Loss of Heterozygosity of APC Gene Detection Exon 11 of Gastric Cancer by Capillary Electrophoresis |
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| DOI：10.3969/j.issn.1004-4957.年份.月份 |
| KeyWord:gastric cancer loss of heterozygosity capillary electrophoresis polyacrylamide gel electrophoresis single strand conformation polymorphism restriction fragment length polymorphism |
| Author | Institution |
| 谢希晖，王荣，谢华，贾正平，张爱梅，孟宪栋，马骏 |
1.兰州大学药学院;2.兰州军区兰州总医院临床药理基地;3.西北师范大学生命科学学院 |
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| Abstract: |
| In this paper，the part base sequences of APC gene on exon 11 were extracted from normal tissue and gastric cancer tissue and amplified by polymerase chain reaction（PCR），which contains some bases easily resulting in loss of heterozygosis（LOH）.Then the amplified samples were denatured at 96 ℃ and digested by restriction enzyme of RsaⅠand detected by capillary electrophoresis（CE）-single strand conformation polymorphism（SSCP），capillary electrophoresis-restriction fragment length polymorphism（RFLP） and polyacrylamide gel electrophoresis（PAGE）-SSCP.The concentration of polyacrylamide gel was selected to be 15%.The analysis of APC gene on exon 11 was performed by CE combined with laser induced fluorescence detector（λex=488 nm，λem=520 nm），by using 3.0% polyethylene oxide（PEO） as sieving medium，1×TBE（pH 8.2） as electrophoresis buffer with a separation voltage of 15 kV at 15 ℃.The sequence of detection ratios for APC gene detected by different methods was as follows:CE-SSCP（30%）>PAGE-SSCP（25%）>CE-RFLP（20%）.The results revealed that the LOH of APC gene in gastric cancer tissue was higher than 10%，and the detection ratio for CE-SSCP was higher than that for PAGE-SSCP and CE-RFLP.Therefore，with the advantages of rapidness and high sensitivity，the CE-SSCP method could lay a good foundation for the convenient,reliable and early diagnosis on gastric cancer. |
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