Increasing drug resistance in human lung cancer cells by mutant-type p53 gene mediated by retrovirus
Tóm tắt
Human mutant-type (mt) p53 cDNA was synthesized and cloned from human lung cancer cell line GL containing mt-p53 gene by using polymerase chain reaction (PCR). It was confirmed that the mt-p53 cDNA contained the complete coding sequence of p53 gene but mutated at codon 245 (G→T) and resulted in glycine to cysteine by sequencing analysis. The retroviral vector pD53M of the mt-p53 was constructed and introduced into the drug-sensitive human lung cancer cells GAO in which p53 gene did not mutate. The transfected GAO cells strongly expressed mutant-type p53 protein by immunohistochemistry, showing that pD53M vector could steadily express in GAO cells. The drug resistance to several anticancer agents of GAO cells infected by pD53M increased in varying degrees, with the highest increase of 4-fold,in vitro andin vivo. By quantitative PCR and flow cytometry (FCM) analyses, the expression of MDR1 gene and the activity of P-glycoprotein (Pgp) did not increase, the expression of MRP gene and the activity of multidrug resistance-related protein (Mrp) increased slightly. These results indicated that the drug resistance associated with mt-p53 gene might be somewhat correlated with MRP/Mrp but not with MDR1/Pgp. It was possible to modify the tumor drug resistance by changing status of p53 gene.
Tài liệu tham khảo
Mitsudomi, T., Minna, J. D., Mutations of 53 gene in human lung cancer cell lines,Oncogene, 1992, 7:171.
Oren, M., Relationship of p53 to the control of apoptotic cell death,Semin. Cancer Biol., 1994, 5:221.
Kerr, J. F. R., Winterfold, C. M., Harmon, B. V., Apoptosis, its significance in cancer and cancer therapy,Cancer, 1994, 73:2013.
Gao, Z. Q., Gao, Z. P., Mutations of p53 and ras genes in human lung cancer cell line GL, inStudy on Basic Medical Sciences (ed. Ge, L. H.) (in Chinese), Beijing: China Personnel Press, 1995, 581–584.
Gao, Z. P., Gao, Z. Q., Coexpression of MDRl and MRP genes in human lung cancer cells, inAdvance of Histochemistry and Cyochemistry, Proceedings of 4th China-Japan Joint Histochemistry & Cytochemistry Symposium (1996), Chongqing: Chongqing Press, 1996, 60–61.
Sambrook, J., Fritsh, E. F., Maniatis, T.,Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, 1989.
Gao, Z. Q., Gao, Z. P., Zhang, X. B., Suppression of human lung cancer by wild-type p5 3,Chin. J. Prev. & Control Chronic Non-communi. Dis. (in Chinese), 1996, 2: 63.
Gao, Z. Q., Gao, Z. P., Zhang, X. B., Inhibitory effect of drug resistance in multidrug-resistant human lung cancer cells by GST-π antisense RNA mediated by retrovirus,Chin. J. Prev. & Control Chronic Non-commumi. Dis. (in Chinese), 1996, 3:110.
Gao, Z.Q., Gao, Z.P., Quantitative analysis of tumor multidrug resistance by PCR,J. Dia. Pathol. (in Chinese), 1996, 3:97.
Kopnin, B. P., Stromskaya, T. P., Kondratov, R. V.et al., Influence of exogenous ras and p53 on p-glycoprotein function in immortalized rodent fibroblasts,Oncol. Res., 1995, 6:299.
Gao, Z. Q., Zheng, J., Wu, B. Q.et al., Study on the biology of xenograft of human lung cancer induced by benzo-(a) pyrene in nude mouse,Chin. J. Pathol. (in Chinese), 1996, 2:106.
Wattel, E., Preudhomme, C., Hecquet, B.et al., P53 mutations are associated with resistance to chemotherapy and short survival in hematologic malignancies,Blood, 1994, 84:3148.
Bardeesy, N., Beckwith, J. B., Pelletier, J., Clonal expansion and attenuated apoptosis in Wilms’ tumors are associated with p53 gene mutations,Cancer Res., 1995, 55:215.
Nabeya, Y., Loganzo, F., Maslak, P.et al., The mutational status of p53 protein in gastric and esophageal cancer cells predicts sensitivity to chemotherapy,Int. J. Cancer, 1995, 64:37.
Preudhomme, C., Dervite, I., Wattel, E.et al., Clinical significance of p53 mutations in newly diagnosed Burkitt’s lymphoma and acute lymphoblastic leukemia,J. Chin. Oncol., 1995, 13:812.
Fujiwara, T., Zhang, W.W., Roth, J. A.et al., Induction of chemotherapy in lung cancer cellsin vim by wild-type p.53 gene,Cancer Res., 1994, 54:2287.
Gao, Z. Q., Gao, Z. P., Advance in multidrug resistance,Chin. J. Pathol. (in Chinese), 1996, 3:185.