China Journal of Oral and Maxillofacial Surgery Vol.9 No.6 November,2011 467 [ ]1672-3244(2011)06-0467-05 DC-CIK 1 2 1 1 (1. 200011; 2. 200031) [ ] : (DC) (CIK) : DC, CIK, DC CIK DC-CIK ; Tca8113 A B C; 24h,A,B DC-CIK,C DC- CIK 4 ; SAS 6.12 t χ 2 :A 7.63d,B 9.5d,C 12d,A C (P=0.0132); 2,A 100%,B 87.5%,C 62.5%,P>0.05;,A,B,C,A B (P>0.05),A C (P=0.036) : DC-CIK Tca8113 [ ] ; ; ; [ ] R739.86 [ ] A The anti-tumor effect of co-cultured DC-CIK from peripheral blood to human tongue SCC cell line Tca8113 in vivo LI Si-yi 1, ZHANG Shang-quan 2, ZHANG Chen-ping 1, CHEN Wan-tao 1. (1.Department of Oral and Maxillofacial Surgery, Ninth People's Hospital, College of Stomatology,Shanghai Jiao Tong University School of Medicine. Shanghai 200011; 2. Institute of Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. Shanghai 200031, China) [Abstract] PURPOSE: To evaluate the anti-tumor effect of co-cultured sensitized dendritic cell (DC) and cytokineinduced killer cell (CIK) separated from peripheral blood to human tongue SCC nude mice cancer model in vivo. METHOD: Both DC and CIK were separated and cultured in vitro from peripheral blood of oral tongue cancer patients. The sensitized DC was co-cultured with CIK. Tca8113 cells were injected into the right oxter of 24 BALB/c nude mice which were divided into control group A, experiment group B and group C. 24 hours later, group A received normal saline injection in one site with Tca 8113 transplantation, group B were injected DC-CIK in opposite side, group C were injected the same dose of DC-CIK in right oxter. The number and size of induced tumors were observed in the next 4 weeks.the data was analyzed with SAS 6.12 software package for student s t test, ANOVA and Chi-square test. RESULTS: The average day of tumor induction was 7.63 days, 9.5 days and 12 days for group A, B and C, respectively, which showed significant difference between group A and C (P=0.0132). Two weeks after Tca8113 injection, 100% of mice in group A developed tumors, 87.5% in B and 62.5% in C developed tumors, which showed no significant difference in 3 groups. The curve of induced tumors size showed that group A developed faster, group C was the slowest with significant difference( P= 0.036). CONCLUSION: The co -cultured sensitized DC -CIK showed a certain anti-tumor effect nude mice with Tca 8113 model in vivo. Supported by the Biomedical Key Research Grant from Science and Technology Commission of Shanghai Muncipality(10DZ1951300). [ ] 2011-06-22; [ ] 2011-09-08 [ ] (10DZ1951300) [ ] (1971-),E-mail:li.siyi@tom.com [ ],Tel:021-23271699-5154,E-mail:zhang.chenping@hotmail.com c 2011
468. DC-CIK LI Si-yi, et al. The anti-tumor effect of co-cultured DC-CIK from peripheral blood to human tongue SCC cell line Tca8113 in vivo [Key words] Tongue; Squamous cell carcinoma; Dendritic cell; Cytokine-induced killer cell;nude mouse China J Oral Maxillofac Surg,2011,9(6):467-471. DC; DC CIK,, DC-CIK;24 Tca811,3 (APC) A B C 24h,A [1-3] (dendritic cell,dc) DC-CIK,C DC-CIK, 1 DC-CIK,48h, [4] T, CIK (cytokine-induced killer cell,cik), 4,CIK CD3 + CD56 +, LAK 1.3.1 DC [9], MHC,CIK [5-6],DC CIK, 1.3.2 CIK CD3 CIK [7-8] CIK IL-12 TNF-α PBMC 1 10 6 DC, DC CIK, 25cm 2 RPMI1640+5% DC-CIK,DC- AB 10mL 1 10 5 /ml IFNγ 1000u/mL,37 5%CO 2 24h CIK Tca8113 PBS, Tca8113, DC-CIK IL-1β IL-2, 100u/ 1 1.1 BALB/c-nu 25, 5~6, 9d CIK DC CIK=1 50 19~23g DC CIK 25cm 2 ;Tca8113 : RPMI1640+5%AB 10mL,IL-1β 100u/mL,IL-2 300u/mL,37 5%CO 2 5d 1.2 Ficoll (Lymphoprep, ); RPMI1640 (Gibo );rhgm-csf rhtnfα rhil-4(peprotech );IFN-γ(,200μL/, FITC CD3,PE ); CD3 ( ) CD56 10μL, IgG1 IgG2 rhil-2( ); AB ),B 3,HE : CD3 ( 5μg/mL) PBS 5mL 25cm 2,4 24h ml 300u/mL,37 5%CO 2 24h 1 10 7 /ml 1.3.3 DC+CIK 9d DC, PBS,RPMI1640+5%AB 9d CIK 14d DC-CIK 0.5mL,PBS,1000rpm PBA, 5 10 5 /ml,4 30min PBS 1mL 1.3 1.3.4 Tca8113 7 25cm 2 DC CIK Tca8113, RPMI1640+5%AB, Tca8113 5mL,37 5%CO 2
China Journal of Oral and Maxillofacial Surgery Vol.9 No.6 November,2011 469 [10] 1.3.5 9.22% ;DC CIK 5d, CD3 + Tca8113 CD56 + 57.62%,CD3 + 97.41% BALB/c 24, 2.1.2, Tca8113 0.2mL, 2 10 6 A B C 3 8, 4 24h 14d DC-CIK Tca8113 ; Tca8113 DC-CIK,,,PBS,PBS,, ; 48h 1 10 8 /ml A HE, 0.3mL,B, DC-CIK 0.2mL, 2 2.2 10 7,C DC-CIK,A, SPF 4 1.4 1.4.1 1),A C 1.4.2 3d 1. A B C Table 1. Days for tumor formation in group A, B and C 3d 1.4.3 14d ( / 100%) 1.4.4, 14d,A B C 3 8 7 5 1.5 ( 2) χ 2, ( < SAS6.12 t 5),3,P=0.083( ),A C χ 2,P<0.05,P=0.20,>0.05, 2.3 2 2.1 2.1.1 CIK CD3 + 9d, 98.41%,CD3 + CD56 + A DC-CIK 6~10d 7.63d;B,C 6,2 9.5d 12d( (P=0.0132),B C (P=0.1902) (d) 1# 2# 3# 4# 5# 6# 7# 8# A 10 8 9 6 6 7 7 8 7.63 B 9 8 6 8 10 10 16 9 9.50 C 10 11 11 23 9 8 12.00 (d) P=0.013(A C ) P=0.190(B C ) A,,C 4,A (184.51±46.13)mm 2,B (160.59±38.36)mm 2,C (95.62±28.53)mm 2 A C,P=0.036, DC-CIK Tca8113 ( 1 2 ) B C 11d A B C 21d 1. Tca8113 11d 21d,A B C Figure 1. The size of tumor in group A, B and C after 11 days and 21 days of Tca8113 injection
470. DC-CIK LI Si-yi, et al. The anti-tumor effect of co-cultured DC-CIK from peripheral blood to human tongue SCC cell line Tca8113 in vivo 2. 14d A B C 3 Table 2. The tumor formation rate of groups in 14 days after tumor cell injection A 0 8 100% B 1 7 87.5% C 3 5 62.5% P=0.20(A,C ) 2.4 3 20.00 0.00 11 14 16 19 23 27 31,A B,C HE,3,,, 2. ( 3) (mm 2 ) 200.00 180.00 160.00 140.00 120.00 100.00 80.00 60.00 40.00 (d) A B C 3 Figure 2. Tumor growth curve of group A, B and C A B C A B C 3 200 400 200 3. A B C HE Figure 3. The gross specimen and HE staining image of transplanted tumors in group A, B and C CIK DC CIK, CIK,, ;,CIK Th1 DC-CIK LAK TIL, T DC-CIK48h, DC, DC-CIK CTL,, BALB/c-nu Tca8113 [11-13] CIK,, LAK CTL 6~12d DC-CIK Tca8113
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