Cyclin D1 antisense oligodexoyneucleotides inhibits growth and enhances chemosensitivity in gastric carcinoma cells.
OBJECTIVE
To examine the effects of cyclin D1 antisense oligodexoyneucleotides (ASODN) on growth and chemosensitivity of gastric carcinoma cell lines SGC7901 and its mechanism.


METHODS
Phosphorothioate modified cyclin D1 ASODN was encapsulated by LipofectAMINE2000 (LF2000) and transfected into cells, the dose-effect curves and growth curves were observed. 5-FU, MTX, CDDP of different concentrations were given after transfecting cells with cyclin D1 ASODN for 24 h the dose-effect responses were observed and IC50s were calculated. The mRNA expression of cyclin D1, thymidylate synthase (TS), thymidine phosphorylase (TP) and dihydrofolate reductase (DHFR) was detected by reverse transcription-PCR (RT-PCR) at 24 h and 48 h after transfection.


RESULTS
Dose-dependent inhibitory effect was caused by cyclin D1 ASODN in SGC7901 cells. Transfecting gastric carcinoma cells with 0.2 micromol/L cyclin D1 ASODN for 24 h could inhibit growth significantly and reduce expression of cyclin D1 mRNA. Cyclin D1 ASODN could increase the chemosensitivity to 5-FU, MTX, CDDP in cells. The IC50s of different chemotherapeutic agents in ASODN plus chemotherapy groups were significantly lower than those in controls. Transfection with cyclin D1 ASODN leaded to an increase in TS and DHFR mRNA and a decrease in TP mRNA as determined by RT-PCR at 24 h, the alterations were more significant at 48 h.


CONCLUSIONS
Cyclin D1 ASODN can decrease mRNA expression of cyclin D1, inhibit growth and enhance the chemosensitivity by changing the expression of enzymes related to metabolism of chemotherapeutic agents in SGC7901 gastric carcinoma cells.
Shuai
Xiao-Ming
XM
Department of Gastroenterologic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China. xmshuai@163.com
Han
Gao-Xiong
GX
Wang
Guo-Bin
GB
Chen
Jun-Hua
JH
D000230:Adenocarcinoma / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002628:Chemoreceptor Cells / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology; D019938:Cyclin D1 / Q000235:genetics* / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D005472:Fluorouracil / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D008727:Methotrexate / Q000494:pharmacology; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013274:Stomach Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics / Q000502:physiology; D013939:Thymidine Phosphorylase / Q000235:genetics / Q000502:physiology; D013940:Thymidylate Synthase / Q000235:genetics / Q000502:physiology; D014162:Transfection
D000970:Antineoplastic Agents; D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D019938:Cyclin D1; D013762:Tetrahydrofolate Dehydrogenase; D013940:Thymidylate Synthase; D013939:Thymidine Phosphorylase; D002945:Cisplatin; D005472:Fluorouracil; D008727:Methotrexate