Identification of spliced variants of the proto-oncogene HDM2 in colorectal cancer.

BACKGROUND

The human double minute 2 (hdm2) oncogene is a negative regulator of the p53 gene. Expression and alternative splicing of the hdm2 gene may contribute to colorectal cancer development or progression. This study aimed to determine the presence and identification of aberrant mRNA transcripts of hdm2 in colorectal cancer tissues and cell lines, and determine the nature of their association with clinicopathological characteristics and survival of patients.

METHODS

A total of 69 colorectal cancer and corresponding normal tissue specimens and 10 colon cancer cell lines were recruited for polymerase chain reaction and DNA sequencing analyses of hdm2 mRNA. Genomic DNA from these tissues and cells was also extracted for p53 gene mutation analysis. The association of hdm2 fragmented transcripts and p53 gene mutation with clinicopathological data was then statistically analyzed.

RESULTS

In 62 cases (89.9%; 62 of 69) of colorectal cancer tissues the full-length hdm2 was amplified, whereas 7 cases had no hdm2 transcripts. Thirty-two of 62 cases (51.6%) and 6 of 10 cell lines (60%) showed at least 1 hdm2 spliced variant. A total of 4 hdm2 splicing variants were found in colorectal cancer tissues and cells, that is, lack of nucleotides between 157 and 292 bp in hdm2/1338, 81 to 901 bp in hdm2/707, 157 to 292, 407 to 505, and 668 to 901 bp in hdm2/1007, and 610 to 883 in hdm2/1200. Of these, hdm2/1338 is a novel hdm2 variant in colorectal cancer. Mutation in p53 was detected in 21 cases (33.8%; 21 of 62). Although there was no association found between expression of hdm2 splicing variants and p53 gene mutations, expression of hdm2 splicing variants was associated with advanced tumor stage (P = .022) and distant metastasis (P = .004) in wild-type p53 cases, and with poor survival of patients (P = .039).

CONCLUSIONS

The data from the current study provide the first evidence that hdm2 mRNA is frequently mutated by alternative splicing in colorectal cancer, and may play a role in colorectal tumorigenesis or cancer progression.