Effects of DC loaded α-GalCer combined with tumor specific cytotoxic T lymphocytes on the growth of transplanted Heps hepatoma in mice
Peng WANG
;
Zhiyong LUAN
;
Junquan LIU
;
Min HANG
;
Jin PAN
;
Nanzheng ZHANG
Journal of International Oncology.2016;43(11):806-811. doi:10.3760/cma.j.issn.1673-422X.2016.11.002
Objective To investigate the effects of dendritic cells (DCs)loading alpha-Galactosylce-ramide (α-GalCer)combined with tumor specific cytotoxic T lymphocytes (CTLs)on the growth of transplanted Heps hepatoma in mice.Methods We induced the augmentation of the DC cells and T lymphocyte derived from the mice bone marrow,and enabled them to be specific CTLs.DC cells loaded α-GalCer in vitro.First we established a Heps liver cancer xenograft model,then divided the model mice into 4 groups by random number table method (n =9):control group (intravenous injection with physiological saline),CTL group,DC loadingα-Galcer group and DC loading α-Galcer combined with CTLs group.After 2 weeks of intervention,we extrac-ted the tumor tissue,weighed the tumor and calculated the inhibition rate of tumor.The expressions of Bax/Bcl-2 cells in groups of transplanted tumor tissues were detected using immunohistochemistry and Western blotting. Results The average tumor weight of CTL group,DC loading α-GalCer group and combined treatment group were (1 .07 ±0.1 5)g,(1 .1 1 ±0.1 7)g,(0.79 ±0.1 4)g,respectively.All of them were lower than that of control group (1 .69 ±0.23)g,with significant differences (t =1 4.1 76,P =0.023;t =1 2.351 ,P =0.034;t =1 8.672,P =0.000).The average tumor weight of combined treatment group was lower than those of the CTL group and DC loading α-GalCer group,with significant differences (t =1 5.236,P =0.01 2;t =1 1 .1 76, P =0.037).Compared to the CTL group (36.69%)and DC loading α-GalCer group (34.32%),the com-bined treatment group had a higher tumor inhibition rate (53.25%;P =0.034,P =0.021 ).Immunohisto-chemical assay showed that the numbers of Bax-positive cells in CTL group,DC loading α-GalCer group and combined treatment group were 35.83 ±0.75,33.67 ±0.82,41 .1 7 ±1 .1 7 respectively,and compared with the control group (21 .67 ±2.1 6),the differences were statistically significant (t =-1 3.789,P =0.002;t =-1 5.1 1 6,P =0.001 ;t =-1 7.452,P =0.000).The numbers of Bax-positive cells in combined treatment group were different with CTL group and DC loading α-GalCer group (t =-7.730,P =0.009;t =-5.872, P =0.01 1 ).The numbers of Bcl-2-positive cells in CTL group,DC loading α-GalCer group and combined treatment group were 30.83 ±0.75,31 .67 ±1 .03,25.00 ±0.89,and compared with the control group (38.67 ±1 .21 ),the differences were statistically significant (t =9.234,P =0.007;t =1 1 .738,P =0.003;t =20.608,P =0.000).The numbers of Bcl-2-positive cells in combined treatment group were different with CTL group and DC loading α-GalCer group (t =1 1 .952,P =0.003;t =1 2.223,P =0.002).Western blot-ting test results showed that the expression levels of Bax in CTL group,DC loading α-GalCer group and com-bined treatment group were 0.46 ±0.01 ,0.42 ±0.03,0.55 ±0.01 ,and compared with the control group (0.31 ±0.02),the differences were statistically significant (t =1 .035,P =0.032;t =1 .1 24,P =0.027;t =1 .425,P =0.01 0).The expression level of Bax in combined treatment group was different with CTL group and DC loading α-GalCer group (t =1 .305,P =0.01 3;t =1 .421 ,P =0.01 0).The positive expressions of Bcl-2 in CTL group,DC loading α-GalCer group and combined treatment group were 0.34 ±0.03,0.33 ± 0.02,0.24 ±0.01 ,and compared with the control group (0.46 ±0.01 ),the differences were statistically sig-nificant (t =-1 .1 23,P =0.025;t =-1 .061 ,P =0.031 ;t =1 .278,P =0.01 4);the positive expression level of Bcl-2 in combined treatment group was different with CTL group and DC loading α-GalCer group (t =1 .1 60,P =0.021 ;t =1 .21 9,P =0.01 5).Conclusion It has synergistic killing effect on transplanted Heps hepatoma in mice using DC loading α-GalCer combined with the tumor specific CTL.
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