[보고서]동물종양모델에서 대장암항원규명을 통한 종양제어기술 개발 및 관련 기전 연구

신정임

보고서 정보

주관연구기관

강원대학교
Kangwon National University

연구책임자

신정임

보고서유형

최종보고서

발행국가

대한민국

언어

한국어

발행년월

2021-06

과제시작연도

2021

주관부처

교육부
Ministry of Education

등록번호

TRKO202100010947

과제고유번호

1345332767

사업명

개인기초연구(교육부)(R&D)

DB 구축일자

2022-04-23

키워드

종양항원.CD8+ T 임파구.면역회피.tumor antigen.anti-PD-L1.IL-12.CD8+ T cell.immune escape.

표 Expression status of PD-L1 on TC-1 and MC38 cells, and the antitumor therapeutic effect of anti-PD-L1 Abs in association with gemcitabine. A) TC-1 cells were stained with PE-conjugated control IgG isotype (thin line) and PE-conjugated anti-PD-L1 Abs (thick line). B,C) Each group of mice (n=5/group) was challenged with TC-1 cells. When the tumor sizes became 2 mm in mean diameter, the mice were injected i.p. with 100 μg of anti-PD-L1 IgGs at 0, 3, 7, 10, and 13 days post-treatment. Tumor sizes were measured over the time points (B). On day 15, the mice were tested for in vivo CTL lytic activity (C). D) Similar experiments to Fig. 1B except that when the tumor sizes become 5 mm in mean diameter, the mice were treated with anti-PD-L1 Abs and gemcitabine. In particular, the mice were injected i.p. with gemcitabine twice a week at a dose of 37.5 mg/g body weight. E) MC38 cells were stained with PE-conjugated control IgG isotype (thin line) and PE-conjugated anti-PD-L1 Abs (thick line). F) Similar experiments to Fig. 1B except with MC38 cells. G) Similar experiments to Fig. 1D except with MC38 cells. *p<0.05 compared to non-treatment. #p<0.05 compared to anti-PD-L1. **p<0.05 compared to gemcitabine+control Ab (by independent t test)
표 Expression status of PD-L1 on B16 cells, and the antitumor therapeutic effect of anti-PD-L1 Abs and the role of immune cell subsets in antitumor immunity. A) B16 cells were stained with PE-conjugated control IgG isotype (thin line) and PE-conjugated anti-PD-L1 Abs (thick line). B) Each group of mice (n=5/group) was challenged with B16 cells. When the tumor sizes became 2 mm in mean diameter, the mice were injected i.p. with 100 μg of anti-PD-L1 Abs at 0, 3, 7, 10, and 13 days post-treatment. The tumor sizes were measured over the time points. The values and bars represent mean tumor sizes and the SD, respectively. C) similar experiments to Fig. 2B except that the mice were injected with anti-PD-L1 Abs at 0, 3, 7, 10, and 13 days post-treatment. The mice were also injected with anti-CD4, anti-CD8, and anti-NK Abs at 0, 3, 7, 10 days post-treatment. *p<0.05 compared to non-treatment. #p<0.05 compared to anti-PD-L1
표 Antitumor memory responses to parental B16 cell challenges in B16 tumor-removed animals under anti-PD-L1 Ab therapy. Each group (n=5/group) of mice was challenged s.c. at the right flank with B16 cells. When the tumor sizes became 2 mm in mean diameter, the mice were injected i.p. with 100 μg of anti-PD-L1 Abs at 0, 3, 7, 10, and 13 days post-treatment. At 15 days post-treatment, tumors (5-6 mm in mean diameter) were surgically removed from mice receiving anti-PD-L1 therapy. After 6 (A) and 3 (B) weeks post-tumor removal, the mice were re-challenged at the left flank with 1 x 106 B16 cells in parallel with age-matched control mice. The tumor sizes were measured over the time points. The values and bars represent mean tumor sizes and the SD, respectively
표 Populations of CD4+ and CD8+ T cells and NK cells in the tumor-draining lymph nodes (TDLNs) and tumor tissues. Each group (n=5/group) of mice was challenged s.c. with B16 cells. When the tumor sizes became 2 mm in mean diameter, the mice were injected i.p. with 100 μg of anti-PD-L1 Abs at 0, 3, 7, 10, and 13 days post-treatment. At 15 days post-treatment, the mice were sacrificed, and the TDLNs and tumor tissues were obtained. Immune cells of the TDLNs and tumor tissues were analyzed for the population of CD4+ and CD8+ T cells, and NK cells
표 Populations of CD4+ and CD8+ T cells, and NK cells in the TC-1 and MC38 tumor tissues. Each group (n=5/group) of mice was challenged s.c. with TC-1 and MC38 cells. After 10 days following tumor cell challenge (tumor sizes; 8~10 mm), the mice were sacrificed, and the tumor tissues were obtained. Immune cells of the tumor tissues were analyzed for the population of CD3+/CD8+ T cells and CD3+/CD4+ T cells, and CD3-/CD49b+ NK cells. *p<0.05 compared to TC-1
표 Immune cell responses to tumor antigens and melanoma class I peptides in tumor-bearing mice under anti-PD-L1 therapy. Each group (n=5/group) of mice was challenged s.c. with B16 cells. (A) When the tumor sizes became 2 mm in mean diameter, the mice were injected i.p. with 100 μg of anti-PD-L1 Abs and control Abs at 0, 3, 7, 10, and 13 days post-treatment. At 15 days post-treatment, the mice were sacrificed and the spleen was obtained. Six x 106 splenocytes per ml were stimulated with 2 x 106 B16 cells, along with TC-1 cells as a control that had been exposed to UV light for 1.5h. Five μg of peptides per ml were also added to the splenocytes. After 2 days stimulation, the cell supernatants were collected for measurement of IFN-γ. Values and bars represent IFN-γ concentrations and the SD. (B) The splenocytes of the B16 tumor bearing mice (tumor size: 10 mm) were stimulated for 1 day with B16 cells and TC-1 cells as described in Fig. 6A. In this case, 1 μM of monensin was added to the cell culture to block the secretion of IFN-γ. The immune cells were stained with anti-IFN-γ, along with each immune cell marker. (C) shows the percentages of CD4+ T cells, CD8+ T cells and CD49b+ NK cells among CD45+ cell populations. (D) shows the percentages of IFN-γ-positive cells among each population of CD4+ and CD8+ T cells, and NK cells. (E) The splenocytes were stimulated for 1 and 2 days with B16 cells and TC-1 cells and the levels of IFN-γ were measured, as described in Fig. 6A, except for no addition of monensin. (F) The splenocytes were stimulated for 2 days with B16 cells and the levels of IFN-γ were measured, as described in Fig. 6A. CD8+ T and NK cell-depleted immune cells were reacted with B16 cells, in parallel with whole immune cells. *p<0.05 compared to TC-1. #p<0.05 compared to non-depletion
표 The levels of HER2 DNA (A,B), mRNA expression (C,D), and protein expression (E) in CT26/HER2, CT26/HER2-A1 and CT26/HER2-A2 cells, along with DNA sequencing schemes (F) and the expression status of HSP90 (G). (A) The tumor cells were lysed for genomic DNA purification. Four hundred nanograms of genomic DNA were tested in a PCR assay for HER2, as described in the Materials and Methods. (B) Four hundred nanograms of genomic DNA were also tested using qRT-PCR assay. Ct values are defined as the number of cycles required for the fluorescent signal to cross the threshold. (C) The tumor cells were treated with TRIzol and then the RNA was isolated, followed by RT-PCR assay. (D) cDNA (generated by RT) was also tested using qRT-PCR assay. The data were used to calculate the levels of HER2 mRNA expression, as described in the Materials and Methods. (E) The tumor cells were lysed in RIPA buffer, and 30 μg samples of the cell lysates were separated by SDS-PAGE and analyzed by Western blot assay against HER2 proteins, as described in the Materials and Methods. (F) shows the whole HER2 gene regions of genomic DNAs and their upstream and downstream regions from CT26/HER2-A1 and –A2 cells that were amplified by PCR using forward and reverse primers (arrows). The PCR products were purified and sequenced. (G) Similar experiments to those shown in Fig. 1E except using anti-HSP90 antibodies for Western blot assay
표 Evaluation of HER2 expression and of Ag-specific immune stimulatory activity in CT26/HER2-A2 cells by treatment with MG132 or chloroquine. (A) CT26/HER2-A2 cells were incubated for 1 day with an increasing concentration of MG132 and chloroquine. The cells were reacted with 2 μl of anti-HER2 sera and then stained with FITC-conjugated anti-mouse IgG for flow cytometry. Thin line; non-treatment, thick line; drug treatment. Anti-HER2 sera were obtained from mice immunized twice with HER2 DNA vaccines. DMFI was calculated as [the MFI values of drug treatment - the MFI values of non-treatment]. (B) CT26/HER2-A2 cells were incubated for 1 day with an increasing concentration of MG132. The tumor cells were lysed in RIPA buffer, and 30 mg samples of the cell lysates were separated by SDS-PAGE and analyzed by Western blot assay. (C) Mice were immunized by IM-EP with HER2 DNA vaccines at 0 and 1 weeks. At 2 weeks, the mice were sacrificed to obtain splenocytes. The splenocytes were incubated for 2 days with CD26/HER2-A2 cells that had been treated for 1 day with 5 μM MG-132 and chloroquine and then exposed to UV light for 3 h prior to immune cell treatment. The cell supernatants were collected for IFN-γ assay. *p<0.05 compared to CT26. **p<0.05 compared to control mice
표 Evaluation of HER2 expression and of Ag-specific immune stimulatory activity in CT26/HER2-A2 cells by treatment with cytotoxic drugs. (A) CT26/HER2-A2 cells were incubated for 1 day with 1 mg/ml of drugs (5-FU, gemcitabine, cisplatin, holoxan, bleomycin, padexol). The tumor cells were lysed in RIPA buffer, and 30 mg samples of the cell lysates were separated by SDS-PAGE and analyzed by Western blot assay. (B) Mice were immunized by IM-EP with HER2 DNA vaccines at 0 and 1 weeks. At 2 weeks, the mice were sacrificed to obtain splenocytes. The splenocytes were incubated for 2 days with CT26/HER2-A2 cells that had been treated for 1 day with 1 μg of each drug per ml and then exposed to UV light for 3 h prior to immune cell treatment. The cell supernatants were collected for IFN-γ assay. *p<0.05 compared to CT26. **p<0.05 compared to control mice
$Evaluation of HER2 mRNA levels in the monosomes and polysomes of CT26/HER2 cells vs. CT26/HER2-A2 cells. (A) CT26/HER2 and CT26/HER2-A2 cells were grown to 80-90% confluency. The cells were harvested to obtain cell lysates, as described in the Materials and Methods. The cells lysates were loaded to sucrose gradients and centrifuged. The OD values of each collected fraction were graphed. The selected fractions containing the monosomes and polysomes of CT26/HER2 and CT26/HER2-A2 cells are indicated by arrows. (B) The selected fractions were treated with TRIzol and RNAs were isolated, followed by RT-PCR, as described in the Materials and Methods$ 표 Evaluation of HER2 mRNA levels in the monosomes and polysomes of CT26/HER2 cells vs. CT26/HER2-A2 cells. (A) CT26/HER2 and CT26/HER2-A2 cells were grown to 80-90% confluency. The cells were harvested to obtain cell lysates, as described in the Materials and Methods. The cells lysates were loaded to sucrose gradients and centrifuged. The OD values of each collected fraction were graphed. The selected fractions containing the monosomes and polysomes of CT26/HER2 and CT26/HER2-A2 cells are indicated by arrows. (B) The selected fractions were treated with TRIzol and RNAs were isolated, followed by RT-PCR, as described in the Materials and Methods
$Preparation of peptide extracts from B16 cells and their fractionation by reversed-phase high performance liquid chromatography (RP-HPLC) and immunogenicity. (A) Peptides were extracted from B16 cells and desalted using the C18 Sep-Pak column, as described in “Materials and method.” The peptides were eluted with 3 ml of 60% acetonitrile (ACN) in water, of which 300 μl was vacuum-dried and then dissolved in 10% DMSO in PBS. The dissolved B16 cell elute was reacted with 6 x 106 splenocytes from mice that had been immunized twice at 2-week intervals with Trp2 peptides plus CpG-ODN. Additionally, 10 μg of Trp2 peptides was dissolved in 3 ml of citrate-phosphate buffer, followed by passage through a C18 Sep-Pak column as described above. This sample was eluted with 3 ml of 60% ACN in water, of which 300 μl was dried and reacted with 6 x 106 splenocytes from Trp2 peptide+CpG-ODN-immunized mice. For the IFN-γ control, 5 μg of Trp2 and E7 peptides were tested. (B) Negative control (E7 peptide), B16 cell elute, and 1 x 106 of B16 and TC-1 cells were reacted with 6 x 106 splenocytes from B16-bearing mice that had been treated with anti-PD-L1. After 2 days of incubation, the cell supernatants were collected for the IFN-γ assay. (C) Dried peptides from 1 ml of elute were dissolved in DMSO and fractionated by RP-HPLC, as described in “Materials and method.” The fractions displaying a high peak at 214 nm were selected and vacuum-dried. Blue line; peaks at 280 nm. (D) The dried fraction samples were re-suspended in 10% DMSO in water and stimulated for 3 days with 6 x 106 splenocytes from B16 tumor-bearing mice receiving anti-PD-L1. The cell supernatants were measured for IFN-γ levels. Arrows indicate the fraction number displaying the highest levels of IFN-γ production among the tested samples. *p<0.05 compared with naïve control. #p<0.05 compared with E7 peptide$ 표 Preparation of peptide extracts from B16 cells and their fractionation by reversed-phase high performance liquid chromatography (RP-HPLC) and immunogenicity. (A) Peptides were extracted from B16 cells and desalted using the C18 Sep-Pak column, as described in “Materials and method.” The peptides were eluted with 3 ml of 60% acetonitrile (ACN) in water, of which 300 μl was vacuum-dried and then dissolved in 10% DMSO in PBS. The dissolved B16 cell elute was reacted with 6 x 106 splenocytes from mice that had been immunized twice at 2-week intervals with Trp2 peptides plus CpG-ODN. Additionally, 10 μg of Trp2 peptides was dissolved in 3 ml of citrate-phosphate buffer, followed by passage through a C18 Sep-Pak column as described above. This sample was eluted with 3 ml of 60% ACN in water, of which 300 μl was dried and reacted with 6 x 106 splenocytes from Trp2 peptide+CpG-ODN-immunized mice. For the IFN-γ control, 5 μg of Trp2 and E7 peptides were tested. (B) Negative control (E7 peptide), B16 cell elute, and 1 x 106 of B16 and TC-1 cells were reacted with 6 x 106 splenocytes from B16-bearing mice that had been treated with anti-PD-L1. After 2 days of incubation, the cell supernatants were collected for the IFN-γ assay. (C) Dried peptides from 1 ml of elute were dissolved in DMSO and fractionated by RP-HPLC, as described in “Materials and method.” The fractions displaying a high peak at 214 nm were selected and vacuum-dried. Blue line; peaks at 280 nm. (D) The dried fraction samples were re-suspended in 10% DMSO in water and stimulated for 3 days with 6 x 106 splenocytes from B16 tumor-bearing mice receiving anti-PD-L1. The cell supernatants were measured for IFN-γ levels. Arrows indicate the fraction number displaying the highest levels of IFN-γ production among the tested samples. *p<0.05 compared with naïve control. #p<0.05 compared with E7 peptide
$Numerous peptide sequences were obtained from the HPLC fractions (B7, B9 and C6) using mass spectrometry. The peptides with relatively high class I binding values in the two class I binding molecule prediction programs, Syfpeithi (http://syfpeithi.de/bin/MHCServer.dll/FindYourMotif.htm) and Rankpep (http://imed.med.ucm.es/Tools/rankpep.html) were synthesized. The values in ( , ) indicate (affinity counts using Syfpeithi, affinity counts using Rankpep)$ 표 Numerous peptide sequences were obtained from the HPLC fractions (B7, B9 and C6) using mass spectrometry. The peptides with relatively high class I binding values in the two class I binding molecule prediction programs, Syfpeithi (http://syfpeithi.de/bin/MHCServer.dll/FindYourMotif.htm) and Rankpep (http://imed.med.ucm.es/Tools/rankpep.html) were synthesized. The values in ( , ) indicate (affinity counts using Syfpeithi, affinity counts using Rankpep)
표 The synthesized peptides were reacted at 1 and 5 μg/ml with 6 x 106 splenocytes from B16-bearing mice that had been treated with anti-PD-L1. After 3 days of incubation, the cell supernatants were collected and measured for the level of IFN-γ. In addition, 3 x 106 of TC-1 and B16 cells were reacted for 3 days with the above splenocytes for measurement of IFN-γ levels
표 The antitumor activity of anti-PD-L1 Ab treatment in combination with combination therapy using Trp2 peptides plus CpG-ODN. (A) Each group of mice (n=5/group) was challenged s.c. with B16 cells. When the tumor sizes became 1.5 mm (A,B) and 3 mm (C) in mean diameter, the mice were injected i.p. with 100 μg of anti-PD-L1 Abs at 0, 3, 7, 10, and 13 days post-treatment. The mice were also treated s.c. with Trp2 peptides plus CpG-ODN at 0 and 7 days post-treatment, as described in the “Material and methods”. The tumor sizes were measured over the time points. The values and bars represent mean tumor sizes (A,C) and %complete tumor regression rates (B), and the SD, respectively. *p<0.05 compared to non-treatment. &p<0.05 compared to anti-PD-L1. #p<0.05 compared to anti-PD-L1 and Trp2+ODN+control Ab
표 A diagram showing the antitumor mechanism mediated by anti-PD-L1 therapy and Trp2 peptide therapy in combination
$The antitumor activity of local IL-12 cDNA (pIL-12) treatment in the MC38 tumor model, and evaluation of tumor cell peptides recognized by CD8+ T cells. (A) Each group of mice (n=5/group) was challenged s.c. with MC38 cells. When the tumor sizes became 5 mm in mean diameter, the mice were injected i.m. with 50 μg of IL-12 cDNA by electroporation. The tumor sizes were measured over the time points. The values and bars represent mean tumor sizes. In the tumor size calculation, one of the 5 mice treated with pIL-12 was excluded as its tumor became uncontrolled. (B) MC38 cells were treated with 3 ml of citrate-phosphate buffer, and the cell suspension was passed through Dried peptides from 1 ml of elute were dissolved in 0.1% TFA and then fractionated by RP-HPLC. The fractions displaying a high peak at 214 nm were selected and vacuum-dried. Blue line; peaks at 280 nm. The dried fraction samples were re-suspended in 10% DMSO in water and stimulated for 3 days with 6 x 106 splenocytes from the MC38 tumor-bearing mice receiving IL-12 cDNA treatment. The cell supernatants were measured for IFN-γ levels. Arrows indicate the fraction number displaying the highest levels of IFN-γ production among the tested samples. (D) The fractions B12 were tested for the presence of 9 amino acid-sized peptide molecules using the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In the peaks, a molecular size of 876, 892, 1018 and 1141 Da were observed$ 표 The antitumor activity of local IL-12 cDNA (pIL-12) treatment in the MC38 tumor model, and evaluation of tumor cell peptides recognized by CD8+ T cells. (A) Each group of mice (n=5/group) was challenged s.c. with MC38 cells. When the tumor sizes became 5 mm in mean diameter, the mice were injected i.m. with 50 μg of IL-12 cDNA by electroporation. The tumor sizes were measured over the time points. The values and bars represent mean tumor sizes. In the tumor size calculation, one of the 5 mice treated with pIL-12 was excluded as its tumor became uncontrolled. (B) MC38 cells were treated with 3 ml of citrate-phosphate buffer, and the cell suspension was passed through Dried peptides from 1 ml of elute were dissolved in 0.1% TFA and then fractionated by RP-HPLC. The fractions displaying a high peak at 214 nm were selected and vacuum-dried. Blue line; peaks at 280 nm. The dried fraction samples were re-suspended in 10% DMSO in water and stimulated for 3 days with 6 x 106 splenocytes from the MC38 tumor-bearing mice receiving IL-12 cDNA treatment. The cell supernatants were measured for IFN-γ levels. Arrows indicate the fraction number displaying the highest levels of IFN-γ production among the tested samples. (D) The fractions B12 were tested for the presence of 9 amino acid-sized peptide molecules using the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In the peaks, a molecular size of 876, 892, 1018 and 1141 Da were observed
표 Evaluation of MC38 tumor cell epitopes recognized by Ag-specific CD8+ T cells in the MC38 tumor model. The previously well-known MC38 CD8+ T cell epitopes (A) and 4 peptides (B) with relatively high binding affinity to MHC class I molecules (by Syfpeithi (http://syfpeithi.de/bin/MHCServer.dll/FindYourMotif.htm) and Rankpep (http://imed.med.ucm.es/Tools/rankpep.html) were synthesized and reacted at 1 and 5 μg/ml with 6 x 106 splenocytes from MC38 tumor-bearing mice that had been treated with IL-12 cDNA by IM-EP. After 3 days of incubation, the cell supernatants were collected and measured for the level of IFN-γ. In addition, 3 x 106 of TC-1 and B16 cells were also incubated for 3 days with the above splenocytes for measurement of IFN-γ levels
표 Decreased expression levels of Bcl-2 were not attributed to IL-6 but were correlated with increased sensitivity to chemotherapeutic drugs in B16-S0-1 cells. (A) B16 and B16-S0-1 cells were lysed in RIPA buffer. Thirty μg of cell lysates were run for SDS-PAGE and Western blot assay using Abs recognizing numerous cell regulatory molecules. The values and bars indicate mean relative density and the SD. (B) Two x 106 of B16 and B16-S0-1 cells per 3 ml were added with 1 μg of anti-IL-6 Abs vs. control Abs and then incubated for 2 days. The cells were tested for Bcl-2 expression levels using Western blot assay. (C,D) B16 and B16-S0-1 cells were treated for 3 days with decreasing doses of cisplatin from 4 μg/ml and gemcitabine from 2 μg/ml, and the cell viability was measured using the trypan blue dye exclusion method. Red-colored arrows indicate ID50. (E,F) B16 and B16-S0-1 cells were treated for 2 days with cisplatin (D, 0.5 μg/ml) and gemcitabine (F, 0.5 μg/ml). The frequency of active caspase-3-positive cells was measured using flow cytometry. *p <0.05 compared to B16
표 B16-S0-1 cells were more resistant to granzyme B-mediated caspase activation than B16 cells. (A) B16 and B16-S0-1 cells were lysed in RIPA buffer. Thirty μg of cell lysates were run for SDS-PAGE and Western blot assay using Abs recognizing numerous caspase and granzyme B-associated molecules. The values and bars indicate mean relative density and the SD. (B,C) B16, B16-S0-1 and B16-S1-1 cells were seeded. The next day, the cells were treated with granzyme B, in accordance with the BioPORTER assay protocol. After 1 day incubation, the cells were collected to measure the frequency of active caspase-3-positive cells (B) cleaved caspase-8-positive cells (C) using flow cytometry. *p <0.05 compared to B16
표 B16-S0-1 cells were more resistant to granzyme B-mediated apoptosis than B16 cells. (A) B16 and B16-S0-1 cells were treated with granzyme B, in accordance with the BioPORTER assay protocol. Thirty μg of cell lysates were run for SDS-PAGE and Western blot assay using Abs recognizing granzyme B-associated intracellular molecules. The values and bars indicate mean relative density and the SD. (B) B16, B16-S0-1 and B16-S1-1 cells were treated with granzyme B as above. The next day, the cells were reacted with Annexin V-FITC and PI for evaluation of apoptosis using flow cytometry. %Apoptotic cells were calculated by counting early (lower right) plus late (upper right) apoptotic cells among total cells. *p <0.05 compared to each of granzyme B-untreated cells. (C) B16, B16-S0-1 and B16-S1-1 cells were treated with FITC control molecules, in accordance with the BioPORTER assay protocol. The next day, the cells were tested for fluorescence levels by flow cytometry. *p <0.05 compared to neg. control (cells without FITC transfection)

과제명(ProjectTitle) :	-
연구책임자(Manager) :	-
과제기간(DetailSeriesProject) :	-
총연구비 (DetailSeriesProject) :	-
키워드(keyword) :	-
과제수행기간(LeadAgency) :	-
연구목표(Goal) :	-
연구내용(Abstract) :	-
기대효과(Effect) :	-

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 제목(한글), 저자명(한글), 발행일자, 전자원문, 초록(한글), 초록(영문) 관리번호, 제목(한글), 제목(영문), 저자명(한글), 저자명(영문), 주관연구기관(한글), 주관연구기관(영문), 발행일자, 총페이지수, 주관부처명, 과제시작일, 보고서번호, 과제종료일, 주제분류, 키워드(한글), 전자원문, 키워드(영문), 입수제어번호, 초록(한글), 초록(영문), 목차
저장형식	Text(ASCII format) Excel format
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증

동물종양모델에서 대장암항원규명을 통한 종양제어기술 개발 및 관련 기전 연구
Determination of colon cancer antigens and development of tumor control measures using animal tumor models 원문보기

초록 ▼

목차 Contents

표/그림 (20)

표/그림 (20)

참고문헌 (25)

연구과제 타임라인

관련 콘텐츠

원문 보기

이 보고서와 함께 이용한 콘텐츠

AI-Helper ※ AI-Helper는 오픈소스 모델을 사용합니다.

선택된 텍스트

연합인증

동물종양모델에서 대장암항원규명을 통한 종양제어기술 개발 및 관련 기전 연구 Determination of colon cancer antigens and development of tumor control measures using animal tumor models 원문보기

초록 ▼

목차 Contents

표/그림 (20) 모든 표/그림 보기

표/그림 (20) 슬라이드로 보기

참고문헌 (25)

연구과제 타임라인

전체(0) 논문(0) 특허(0) 보고서(0)

전체(0) 논문(0) 특허(0) 보고서(0)

관련 콘텐츠

원문 보기

이 보고서와 함께 이용한 콘텐츠

AI-Helper ※ AI-Helper는 오픈소스 모델을 사용합니다.

선택된 텍스트

동물종양모델에서 대장암항원규명을 통한 종양제어기술 개발 및 관련 기전 연구
Determination of colon cancer antigens and development of tumor control measures using animal tumor models 원문보기

표/그림 (20)

표/그림 (20)