Antisense oligonucleotide reverses topotecan-resistant ovarian cancer cells

• Published in: Ai zheng = Aizheng = Chinese journal of cancer Vol. 22; no. 12; p. 1296
• Main Authors: Jia, Ping, Wu, Shao-Bo, Xu, Qian, Wu, Ming-Fu, Gao, Qing-Lei, Liao, Guo-Ning, Lu, Yun-Ping, Ma, Ding
• Format: Journal Article
• Language: Chinese
• Published: China 01.12.2003
• Subjects: Antineoplastic Agents - pharmacology; ATP Binding Cassette Transporter, Sub-Family G, Member 2; ATP-Binding Cassette Transporters - antagonists & inhibitors; ATP-Binding Cassette Transporters - genetics; Drug Interactions; Drug Resistance, Neoplasm - drug effects; Drug Screening Assays, Antitumor; Female; Humans; Neoplasm Proteins - antagonists & inhibitors; Neoplasm Proteins - genetics; Oligonucleotides, Antisense - pharmacology; Ovarian Neoplasms - pathology; RNA, Messenger - biosynthesis; RNA, Messenger - drug effects; Topotecan - pharmacology; Transfection; Tumor Cells, Cultured

Breast cancer resistance protein (BCRP) was overexpressed in topotecan (TPT)-selected human ovarian cancer cell line A2780/TPT, strongly suggesting BCRP to be responsible for the drug-resistance of ovarian cancer. The current study was designed to investigate the reversal effect of BCRP antisense oligonucleotide (ASODN) on topotecan- resistant A2780/TPT cells. The antisense-phosphorothioate oligonucleotide including the translation initiation site of BCRP mRNA was artificially synthesized, and the sense oligonucleotide (SODN) corresponding to the ASODN was also synthesized as control. Lipofect-2000 (LF) was used for the transfer of either ASODN or SODN into A2780/TPT cells. The changes of BCRP mRNA expression, intracellular fluorescence intensity of rhodamine and resistance index to topotecan of in vitro transfected A2780/TPT cells were detected respectively by reverse transcription-polymerase chain reaction (RT-PCR),flow cytometry (FCM),and methyl thiazolyl tetrazolium (MTT) assay. The transfer of ASODN/LF i