[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.
Li
Lei
L
Department of Obstetrics and Gynecolology, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing 100730, China.
Xiang
Yang
Y
Tian
Fang
F
Yang
Xiuyii
X
Ren
Tong
T
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