[Transfection of MDR1-mRNA into human mononuclear cells to improve their resistance to anticancer agents, an in vitro study].
OBJECTIVE
To investigate the expression and function of P-glycoprotein (P-gp), an efflux pump encoded by multidrug resistance complementary DNA (MDR1), in human mononuclear cells (MNCs) transfected with MDR1 mRNA.


METHODS
Two kinds of human MDR1 mRNA, pT7TS-MDR1 and pGEM5Zf (+)-MDR1 with difference in the 5' and 3' untranslated regions only, were constructed and modified and then were transfected into human MNCs rich in hematopoietic progenitor cells from a patient with small cell lung cancer. The expression efficiency and pump function of P-gp were measured with FACS. Rhodomine 123 efflux test was used to examine the function of P-gp. Un- transfected mononuclear cells from the same person were used as controls.


RESULTS
The expression of P-gp 12 hours after transfection was 2.16 % in control group, 8.94% in pGEM5Zf (+)-MDR1 group, and 19.14% in pT7TS-MDR1 group. The expression of P-gp 72 hours after transfection was 2.12% in control group, 6.12% in pGEM5Zf(+)-MDR1 group, and 10.71% in pT7TS-MDR1 group, significantly higher in the third group. Rhodamine-123 efflux test showed that the efflux-pump function of P-gp 12 hours after the trransfection was 19.20% in control group, 25.59% in GEM5Zf (+)- MDR1 group, and 35.02% in pT7TS-MDR1 group (all P < 0.01). The expression and presence of P-gp by transfected cells lasted at least 72 hours.


CONCLUSIONS
Transfection of human mononuclear cells with MDR1 mRNA significantly increases their resistance to anticancer agents.
Department of Obstetrics and Gynecolology, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing 100730, China.
Lei
L
Li
Yang
Y
Xiang
Fang
F
Tian
Xiuyii
X
Yang
Tong
T
Ren
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics*; D000970:Antineoplastic Agents / Q000494:pharmacology; D018288:Carcinoma, Small Cell; D019008:Drug Resistance, Neoplasm; D015870:Gene Expression; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008175:Lung Neoplasms; D012333:RNA, Messenger; D014162:Transfection; D014407:Tumor Cells, Cultured
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000970:Antineoplastic Agents; D012333:RNA, Messenger