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Inter‐ alu PCR detects high frequency of genetic alterations in glioma cells exposed to sub‐lethal cisplatin
Author(s) -
Srivastava Tapasya,
Seth Anandita,
Datta Kamal,
Chosdol Kunzang,
Chattopadhyay Parthaprasad,
Sinha Subrata
Publication year - 2005
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/ijc.21057
Subject(s) - alu element , biology , rapd , primer (cosmetics) , cisplatin , microbiology and biotechnology , glioma , dna , polymerase chain reaction , genome instability , genetics , dna damage , genome , gene , chemotherapy , human genome , chemistry , population , medicine , environmental health , organic chemistry , genetic diversity
Increased genomic instability contributes to higher frequency of secondary drug resistance and neoplastic progression in tumors as well as in cells exposed to sub‐lethal concentrations of chemotherapeutic agents. We have used PCR based DNA fingerprinting techniques of randomly amplified polymorphic DNA (RAPD) and inter‐ alu PCR to study this phenomenon in the tumor genome. The choice of the primer, either random (for RAPD) or specific (inter‐ alu PCR) can determine the nature of alterations being assessed. We have compared the inter‐ alu PCR and RAPD profiles of U87MG glioblastoma cells exposed to sequentially increasing low doses of cisplatin for 24 passages to that of untreated controls. Inter‐ alu PCR, with 2 primers, demonstrated a number of alterations in the treated cells, in the form of loss / gain and changes in the intensity of bands. No changes were observed by RAPD analysis with 5 primers, however, indicating a preferential increase in the alu mediated recombination frequency in the treated cells ( p = 1.866 × 10 −4 ). The number of changes observed with respect to the corresponding leucocyte DNA in the inter‐ alu PCR profile of 26 primary tumors (Grade II = 13; Grade IV = 13), resected before chemotherapy, for the 2 inter‐ alu primers was very small. We present a novel application of the inter‐ alu PCR in detecting alterations in long term cultured cells at low dose exposure to a chemotherapeutic agent. Our results suggest that alu mediated recombination may be important in cells exposed to sub‐lethal doses of cisplatin but not in the genesis of primary glioma. © 2005 Wiley‐Liss, Inc.