中华消化外科杂志
中華消化外科雜誌
중화소화외과잡지
CHINESE JOURNAL OF DIGESTIVE SURGERY
2010年
6期
426-429
,共4页
小肠肿瘤%恶性黑色素瘤%肝转移%动物模型%树突状细胞%杀伤细胞%免疫治疗
小腸腫瘤%噁性黑色素瘤%肝轉移%動物模型%樹突狀細胞%殺傷細胞%免疫治療
소장종류%악성흑색소류%간전이%동물모형%수돌상세포%살상세포%면역치료
Neoplasms of small intestine%Malignant melanoma%Liver metastasis%Animal model%Dendritic cells%Killer cells%Immunotherapy
目的 探讨树突状细胞(DC)与同源细胞因子诱导的杀伤细胞(CIK)共培养后对小肠恶性黑色素瘤生长和肝转移的抑制作用.方法 提取健康献血者、小肠恶性黑色素瘤患者外周血单核细胞,常规诱导出DC细胞与CIK细胞后,按照1∶5比例共培养14 d获得DC-CIK细胞.取原发性小肠恶性黑色素瘤手术切除的肝转移灶新鲜瘤组织块植入裸鼠小肠黏膜层,建立小肠恶性黑色素瘤肝转移模型.将56只裸鼠原位移植人小肠恶性黑色素瘤72 h后,分成7组,每组8只:(1)对照组;(2)丝裂霉素组;(3)健康人CIK细胞组;(4)健康人DC-CIK细胞组;(5)小肠恶性黑色素瘤自体CIK细胞组;(6)小肠恶性黑色素瘤自体DC-CIK细胞组;(7)小肠恶性黑色素瘤自体DC-CIK细胞+丝裂霉素组.对照组裸鼠注射生理盐水,其余各组裸鼠尾静脉注射相应的效应细胞,每只0.3 ml,细胞数为6×107个,丝裂霉素注射剂量为400 μg,每天1次,连续28 d.停药后72 h处死动物,切取肿瘤称质量,并检查肿瘤生长及肝转移情况.多组比较采用单因素方差分析,两两比较采用LSD法,率的比较采用x2检验.结果 成功建立小肠恶性黑色素瘤肝转移模型(HSIM-0603).对照组、丝裂霉素组、健康人CIK细胞组和DC-CIK细胞组、小肠恶性黑色素瘤患者自体CIK细胞组和DC-CIK细胞组及DC-CIK细胞+丝裂霉素组的肿瘤质量分别为(1.18±0.17)、(0.71±0.06)、(0.68±0.15)、(0.43±0.08)、(0.67±0.07)、(0.37±0.08)、(0.20±0.05)g;抑瘤率分别为0、39.82%、42.37%、63.56%、43.22%、68.64%、83.05%;肝转移分别为8、8、5、4、4、3、2只,各组比较,差异有统计学意义(F=152.300,x2=200.538,94.325,P＜0.05).结论 小肠恶性黑色素瘤肝转移模型可用于小肠恶性黑色素瘤的发病机制、侵袭、转移及治疗的研究,DC-CIK细胞治疗能抑制小肠恶性黑色素瘤的生长和肝转移.
目的 探討樹突狀細胞(DC)與同源細胞因子誘導的殺傷細胞(CIK)共培養後對小腸噁性黑色素瘤生長和肝轉移的抑製作用.方法 提取健康獻血者、小腸噁性黑色素瘤患者外週血單覈細胞,常規誘導齣DC細胞與CIK細胞後,按照1∶5比例共培養14 d穫得DC-CIK細胞.取原髮性小腸噁性黑色素瘤手術切除的肝轉移竈新鮮瘤組織塊植入裸鼠小腸黏膜層,建立小腸噁性黑色素瘤肝轉移模型.將56隻裸鼠原位移植人小腸噁性黑色素瘤72 h後,分成7組,每組8隻:(1)對照組;(2)絲裂黴素組;(3)健康人CIK細胞組;(4)健康人DC-CIK細胞組;(5)小腸噁性黑色素瘤自體CIK細胞組;(6)小腸噁性黑色素瘤自體DC-CIK細胞組;(7)小腸噁性黑色素瘤自體DC-CIK細胞+絲裂黴素組.對照組裸鼠註射生理鹽水,其餘各組裸鼠尾靜脈註射相應的效應細胞,每隻0.3 ml,細胞數為6×107箇,絲裂黴素註射劑量為400 μg,每天1次,連續28 d.停藥後72 h處死動物,切取腫瘤稱質量,併檢查腫瘤生長及肝轉移情況.多組比較採用單因素方差分析,兩兩比較採用LSD法,率的比較採用x2檢驗.結果 成功建立小腸噁性黑色素瘤肝轉移模型(HSIM-0603).對照組、絲裂黴素組、健康人CIK細胞組和DC-CIK細胞組、小腸噁性黑色素瘤患者自體CIK細胞組和DC-CIK細胞組及DC-CIK細胞+絲裂黴素組的腫瘤質量分彆為(1.18±0.17)、(0.71±0.06)、(0.68±0.15)、(0.43±0.08)、(0.67±0.07)、(0.37±0.08)、(0.20±0.05)g;抑瘤率分彆為0、39.82%、42.37%、63.56%、43.22%、68.64%、83.05%;肝轉移分彆為8、8、5、4、4、3、2隻,各組比較,差異有統計學意義(F=152.300,x2=200.538,94.325,P＜0.05).結論 小腸噁性黑色素瘤肝轉移模型可用于小腸噁性黑色素瘤的髮病機製、侵襲、轉移及治療的研究,DC-CIK細胞治療能抑製小腸噁性黑色素瘤的生長和肝轉移.
목적 탐토수돌상세포(DC)여동원세포인자유도적살상세포(CIK)공배양후대소장악성흑색소류생장화간전이적억제작용.방법 제취건강헌혈자、소장악성흑색소류환자외주혈단핵세포,상규유도출DC세포여CIK세포후,안조1∶5비례공배양14 d획득DC-CIK세포.취원발성소장악성흑색소류수술절제적간전이조신선류조직괴식입라서소장점막층,건립소장악성흑색소류간전이모형.장56지라서원위이식인소장악성흑색소류72 h후,분성7조,매조8지:(1)대조조;(2)사렬매소조;(3)건강인CIK세포조;(4)건강인DC-CIK세포조;(5)소장악성흑색소류자체CIK세포조;(6)소장악성흑색소류자체DC-CIK세포조;(7)소장악성흑색소류자체DC-CIK세포+사렬매소조.대조조라서주사생리염수,기여각조라서미정맥주사상응적효응세포,매지0.3 ml,세포수위6×107개,사렬매소주사제량위400 μg,매천1차,련속28 d.정약후72 h처사동물,절취종류칭질량,병검사종류생장급간전이정황.다조비교채용단인소방차분석,량량비교채용LSD법,솔적비교채용x2검험.결과 성공건립소장악성흑색소류간전이모형(HSIM-0603).대조조、사렬매소조、건강인CIK세포조화DC-CIK세포조、소장악성흑색소류환자자체CIK세포조화DC-CIK세포조급DC-CIK세포+사렬매소조적종류질량분별위(1.18±0.17)、(0.71±0.06)、(0.68±0.15)、(0.43±0.08)、(0.67±0.07)、(0.37±0.08)、(0.20±0.05)g;억류솔분별위0、39.82%、42.37%、63.56%、43.22%、68.64%、83.05%;간전이분별위8、8、5、4、4、3、2지,각조비교,차이유통계학의의(F=152.300,x2=200.538,94.325,P＜0.05).결론 소장악성흑색소류간전이모형가용우소장악성흑색소류적발병궤제、침습、전이급치료적연구,DC-CIK세포치료능억제소장악성흑색소류적생장화간전이.
Objective To investigate the inhibitory effect of dendritic cells (DCs) and homologous cytokine-induced killer cells (CIKs) on the growth and liver metastasis of human primary malignant melanoma of the small intestine. Methods A histologically intact liver metastasis fragment derived from a surgical specimen of patients with primary malignant melanoma of the small intestine was implanted in the mucous layer of small intestine in nude mice to construct the liver metastasis model of the malignant melanoma. Peripheral blood mononuclear cells from healthy donors and patients with malignant melanoma of the small intestine were isolated. DCs and CIKs were separately produced according to corresponding culture method, and the DC-CIKs were harvested after coculorthotopically implanted in 56 nude mice, and 72 hours later, all the mice were equally divided into normal saline group (0.3 ml), mitomycin treatment group (400 μg/0.3 ml), healthy donor CIK group (6 × 107/0.3 ml),healthy donor DC-CIK group (3 × 107/0.3 ml), autologous CIK treatment group (6 × 107/0.3 ml), autologous DC-CIK group (6 × 107/0. 3 ml) and DC-CIK (6 × 107/0.3 ml) + mitomycin (400 μg/0. 3 ml) treatment group. Mice in the experimental groups were administered the agents once daily by vena caudalis injection for 28 days. Mice were sacrificed 72 hours after treatment, and the orthotopic xenografts were removed for weighing,and the liver metastases were simultaneously evaluated by macroscopy and microscopy. All data were analyzed using the analysis of variance, LSD test and chi-square test. Results A liver metastatic model of human primary malignant melanoma of the small intestine ( HSIM-0603 ) was established. Of the control group, healthy donor CIK group, healthy donor DC-CIK group, autologous CIK treatment group, autologous DC-CIK group and DC-CIK +mitomycin group, the tumor weights were ( 1.18 ± 0. 17), (0.71 ± 0.06), (0.68 ± 0. 15 ), (0. 43 ± 0.08 ),(0.67 ± 0.07 ), (0.37 ± 0.08 ) and (0.20 ± 0.05 ) g, respectively; the tumor inhibition rates were 0, 39.82%,42.37%, 65.56%, 43.22%, 68.64% and 83.05%, respectively; liver metastases were detected in 8, 8, 5,4, 4, 3, 2 rats, respectively, in the above mentioned groups. There were significant differences in the tumor weight, tumor inhibition rate and liver metastasis rate among the seven groups (F = 152. 300, x2= 200. 538,94. 325, P ＜ 0.05 ). Conclusions The liver metastatic model could be applied to the studies on the pathogenesis,invasion, metastasis and treatment of human primary malignant melanoma of the small intestine. DC-CIK has the potential in inhibiting the growth and liver metastasis of the malignant melanoma of the small intestine.
