Effect of survivin antisense mRNA transfection on the growth and chemotherapy sensitivity of lymphoma cells.
- Author:
Xia GU
1
;
Han-Liang LIN
;
Jian-Yong SHAO
;
Meng ZHANG
;
You-Kai ZHU
;
Hui-Zhen LIANG
;
Yi-Hui MA
Author Information
- Publication Type:Journal Article
- MeSH: Antimetabolites, Antineoplastic; pharmacology; Antineoplastic Agents, Alkylating; pharmacology; Apoptosis; drug effects; Cell Proliferation; drug effects; Cyclophosphamide; pharmacology; Humans; Inhibitor of Apoptosis Proteins; Jurkat Cells; cytology; metabolism; K562 Cells; cytology; metabolism; Lymphoma; pathology; Methotrexate; pharmacology; Microtubule-Associated Proteins; biosynthesis; genetics; Neoplasm Proteins; biosynthesis; genetics; Plasmids; RNA, Antisense; RNA, Messenger; biosynthesis; genetics; Transfection
- From: Chinese Journal of Pathology 2005;34(11):737-741
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of transfecting survivin antisense mRNA on growth and chemotherapy sensitivity of lymphoma cells.
METHODSEukaryotic 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.
RESULTSCompared 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.
CONCLUSIONSThe 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.