DC-CIK. Tca8113, [ ] (2011) CIK, Tca8113. Tca8113 DC CIK DC-CIK ; t (P=0.0132); , SAS Tca8113

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1 China Journal of Oral and Maxillofacial Surgery Vol.9 No.6 November, [ ] (2011) DC-CIK ( ; ) [ ] : (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 [ ] ; ; ; [ ] R [ ] 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 ; 2. Institute of Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. Shanghai , 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(10DZ ). [ ] ; [ ] [ ] (10DZ ) [ ] (1971-), li.siyi@tom.com [ ],Tel: , zhang.chenping@hotmail.com c 2011

2 468. 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): 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 DC [9], MHC,CIK [5-6],DC CIK, CIK CD3 CIK [7-8] CIK IL-12 TNF-α PBMC DC, DC CIK, 25cm 2 RPMI1640+5% DC-CIK,DC- AB 10mL /ml IFNγ 1000u/mL,37 5%CO 2 24h CIK Tca8113 PBS, Tca8113, DC-CIK IL-1β IL-2, 100u/ BALB/c-nu 25, 5~6, 9d CIK DC CIK= ~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 /ml DC+CIK 9d DC, PBS,RPMI1640+5%AB 9d CIK 14d DC-CIK 0.5mL,PBS,1000rpm PBA, /ml,4 30min PBS 1mL Tca cm 2 DC CIK Tca8113, RPMI1640+5%AB, Tca8113 5mL,37 5%CO 2

3 China Journal of Oral and Maxillofacial Surgery Vol.9 No.6 November, [10] % ;DC CIK 5d, CD3 + Tca8113 CD %,CD % BALB/c 24, 2.1.2, Tca mL, A B C 3 8, 4 24h 14d DC-CIK Tca8113 ; Tca8113 DC-CIK,,,PBS,PBS,, ; 48h /ml A HE, 0.3mL,B, DC-CIK 0.2mL, ,C DC-CIK,A, SPF ),A C d 1. A B C Table 1. Days for tumor formation in group A, B and C 3d d ( / 100%) 1.4.4, 14d,A B C ( 2) χ 2, ( < SAS6.12 t 5),3,P=0.083( ),A C χ 2,P<0.05,P=0.20,>0.05, 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 B C (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. Tca d 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

4 470. 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 % B % C % P=0.20(A,C ) ,A B,C HE,3,,, 2. ( 3) (mm 2 ) (d) A B C 3 Figure 2. Tumor growth curve of group A, B and C A B C A B C 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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