Figure 1 | Oncogene

Figure 1

From: PIK3CA gene is frequently mutated in breast carcinomas and hepatocellular carcinomas

Figure 1

PCR-SSCP of PIK3CA gene in human cancers. Genomic DNA each from normal cells or tumor cells from the same patients was amplified with one primer pair covering exon 9 and with two primer pairs covering exon 20 of PIK3CA gene. Radioisotope was incorporated into the PCR products for detection by autoradiogram. The PCR reaction mixture was denatured for 1 min at 94°C and incubated for 30 cycles. Other procedures of PCR and SSCP analysis were performed as described previously (Kim et al., 2003). After SSCP, DNAs showing mobility shifts were cut out from the dried gel, and re-amplified for 30 cycles using the same primer sets. Sequencing of the PCR products was carried out using the cyclic sequencing kit (Perkin-Elmer, Foster City, CA, USA). Representative data of SSCP (a, b) and DNA sequencing analysis (c, d) of PIK3CA DNA from tumors (Lane T) and normal tissues (Lane N). (a, b) SSCPs of DNA from the tumors (T) show wild-type bands and additional aberrant bands (arrows) as compared to SSCP from corresponding normal cells (N). (c) Sequencing analysis from the aberrant band in (a). There is an A to G transition at nucleotide 3140 of PIK3CA (arrows) in tumor tissue as compared to normal tissue. (d) Sequencing analysis from the aberrant band in (b). There is an insertion of A at the nucleotide between nucleotides 2004 and 2005 of PIK3CA (arrows) in tumor tissue as compared to normal tissue. Numbering of cDNA of PIK3CA was carried out with respect to the ATG start codon (GenBank)

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