论文部分内容阅读
AIM:To isolate and clone the vincristine-resistine-relatedgenes in gastric cancer SGC7901 cell line and to clarify themultidrug-resistant molecular mechanism of gastric cancercells.METHODS:The modified differential-display polymerasechain reaction(DD-PCR)was used to examine thedifferences in the mRNA composition of Vincristine-resistantgastric cancer SGC 7901 cells(SGC7901/VCR),induced byvincristine sulfate versus SGC7901cells.The differentiallyexpressed cDNA fragments were confirmed byreverseNorthern analysis,sequencing,BLAST analysis andNorthern bolt analysis.RESULTS:DD-PCR identified that 54 cDNA fragments werepreferentially expressed in SGC 7901/VCR cells.When thesecDNA fragments were analyzed by reverseNorthern blot,20were reproducibly expressed at a high level in SGC7901/VCR.Sequencing and BLAST analysis revealed that sevenof the genes were known genes:ADP-ribosylation factor 4,cytochrome oxidase subunit Ⅱ,Ss-A/Ro ribonucleoprteinautoantigen 00kd subunit,ribosomal protein S13,galaectin-8 gene,oligophrenin 1 mRNA,and ribosomal protein L23mRNA;and thirteen of the genes were unknown genes.Thelength and abundance of the four unknown genes mRNAwere further confirmed by Northern blot analysis.CONCLUSION:The twenty differential known and unknowngenes may be related to the vincristine-resistant mechanismin human gastric cancer SGC7901 cell line.
AIM: To isolate and clone the vincristine-resistine-relatedgenes in gastric cancer SGC7901 cell line and to clarify themultidrug-resistant molecular mechanism of gastric cancer cells. METHODS: The modified differential-display polymerase chain reaction (DD-PCR) was used to examine the differencies in the mRNA composition of Vincristine-resistant gastric cancer SGC 7901 cells (SGC7901 / VCR) induced by vincristine sulfate versus SGC7901 cells. The differentially expressed cDNA fragments were confirmed by reverse Northern analysis, sequencing, BLAST analysis and Northern bolt analysis. RESULTS: DD-PCR identified that 54 cDNA fragments were screened in a high level in SGC7901 / VCR.Sequencing and BLAST analysis revealed that the seven of the genes were known genes: ADP-ribosylation factor 4, cytochrome oxidase subunit II, Ss-A / Ro ribonucleoproteinautoantigen 00kd subunit, ribosomal protein S13, g alactin-8 gene, oligophrenin 1 mRNA, and ribosomal protein L23 mRNA; and thirteen of the genes were unknown genes. Thelength and abundance of the four unknown genes mRNAwere further confirmed by Northern blot analysis.CONCLUSION: The twenty differential known and unknown genes may be related to the vincristine-resistant mechanismin human gastric cancer SGC7901 cell line.