[Effect of survivin antisense mRNA transfection on the growth and chemotherapy sensitivity of lymphoma cells].
OBJECTIVE
To study the effect of transfecting survivin antisense mRNA on growth and chemotherapy sensitivity of lymphoma cells.


METHODS
Eukaryotic expression plasmid pcDNA3. 1-antisense (As) survivin was constructed and transfected into Jurkat T lymphoblastic lymphoma cell lines with high expression survivin mRNA by use of lipofectmine gene transfer technique. Expression of survivin mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemical and Western blot. The effect of transfecting survivin antisense mRNA on the growth of Jurkat cell lines was monitored by population doubling time (PDT) and Apoptotic indexes (AI). The morphologic features were observed in transfected cells by light and electric microscopes. MTT assay was used to analyze the response of transfected cells to CTX and MTX.


RESULTS
Compared with the control cells, the expression of survivin mRNA and protein were reduced after transfected pcDNA3. 1-Assurvivin 48 h, 5 w and 6 w, PDT (52 h) was prolonged. Apoptotic indexes were higher in transfected antisense survivin mRNA cells [20.2% (48 h)], 6.2% (5 w) and 6.8% (6 w) than control ones [2.1%, 1.3% (48 h)] and [1.3% (5 w) and 1.0% (6 w)]. The cells grow slowly and the dead cells increase and some swelling and apoptotic cells were observed in transfected pcDNA3. 1-Assurvivin groups by invert, light and electric microscopes. The Jurkat cell line of transfected pcDNA3. 1-Assurvivin had higher sensitivity to CTX and MTX. The rate of inhibition was higher in transfected group. There is a significant difference between the transfected group and untransfected one, P < 0.05.


CONCLUSIONS
The result indicated that survivin gene was very important for growth of Jurkat cells. To inhibit the expression of survivin will be significant in therapy of T lymphoblastic lymphoma. Survivin gene might be a target of therapy.
Department of Pathology, Basic Medical College, Sun Yat-sen University, Guangzhou 510089, China.
Xia
X
Gu
Han-Liang
HL
Lin
Jian-Yong
JY
Shao
Meng
M
Zhang
You-Kai
YK
Zhu
Hui-Zhen
HZ
Liang
Yi-Hui
YH
Ma
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D003520:Cyclophosphamide / Q000494:pharmacology*; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D019169:Jurkat Cells / Q000166:cytology / Q000378:metabolism; D020014:K562 Cells / Q000166:cytology / Q000378:metabolism; D008223:Lymphoma / Q000473:pathology; D008727:Methotrexate / Q000494:pharmacology*; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis* / Q000235:genetics; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D000077022:Survivin; D014162:Transfection
D000964:Antimetabolites, Antineoplastic; D018906:Antineoplastic Agents, Alkylating; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D000077022:Survivin; D003520:Cyclophosphamide; D008727:Methotrexate