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Growth inhibition of human pancreatic cancer cell lines by anti‐sense oligonucleotides specific to mutated K‐ ras genes
Author(s) -
Kita Keiichiro,
Saito, Seiji,
Morioka Cintia Y.,
Watanabe Akiharu
Publication year - 1999
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19990209)80:4<553::aid-ijc12>3.0.co;2-6
Subject(s) - point mutation , microbiology and biotechnology , pancreatic cancer , carcinogenesis , sense (electronics) , mutation , transfection , oligonucleotide , biology , cell culture , cancer research , cell growth , cancer , gene , genetics , chemistry
About 90% of human pancreatic cancers carry K‐ ras point mutation, which may play an important role in tumorigenesis. We investigated the inhibitory effects of anti‐sense oligonucleotides targeting K‐ ras point mutation on the growth of cultured human pancreatic cancer cells. Eight human pancreatic cancer cell lines were screened for K‐ ras codon 12 point mutations by PCR‐RFLP analysis and direct sequencing. Then, 3 cell lines with the major types of K‐ ras point mutation, i.e., HuP‐T1, HuP‐T3 and PANC‐1, and 1 without mutation, BxPC‐3, were used for the experiments. Seventeen mer anti‐sense oligonucleotides were designed, targeting the point mutation of K‐ ras codon 12, and transfected into the cells by the liposome‐mediated method. Cell‐growth activities were estimated by MTT assay. Levels of K‐ ras mRNA expression were determined using quantitative RT‐PCR, and K‐ ras p21 protein synthesis was evaluated with Western blotting. Mutation‐matched anti‐sense oligonucleotides effectively inhibited the growth of these pancreatic cancer cell lines, except for BxPC‐3, by suppressing K‐ ras mRNA expression and K‐ ras p21 protein synthesis. Moreover, mutation‐matched anti‐sense oligonucleotides showed stronger anti‐proliferative effects than did mutation‐mismatched ones. Our results suggest that anti‐sense therapy specific to point mutations of K‐ ras mRNA is a practical approach to selective suppression of tumor growth, with little effect on normal cells. Int. J. Cancer 80:553–558, 1999. © 1999 Wiley‐Liss, Inc.