CAJ | 학술논문

目的：探讨不同刺激因子对胃癌中 CXCL9和 CXCL10表达的影响，及不同血样中免疫细胞经不同方式活化后，其膜表面 CXCR3的表达情况，建立并评价 CXCR3和 CXCL9、CXCL10介导的靶向抗胃癌免疫治疗新技术。方法：采用 IFN －γ、TNF －α、Poly（I∶C）、LPS 和 PHA 刺激胃癌细胞（BGC －823、SGC －7901及 MKN－28），ELISA 法测定肿瘤细胞分泌 CXCL9和 CXCL10的情况。将免疫细胞分别活化为 CIK 和 CAPRI，流式细胞术检测活化前后免疫细胞表面 CXCR3的表达情况。四甲基偶氮唑蓝（MTT）法评价刺激因子[IFN －γ、TNF －α、Poly（I∶C）、LPS 和 PHA]本身对肿瘤细胞增殖的影响。Transwell 趋化实验比较趋化因子 CXCL9和CXCL10及其受体 CXCR3的表达差异对免疫细胞靶向归巢至肿瘤的数量的影响。结果：胃癌细胞株基础状态无明显 CXCL9和 CXCL10的表达，IFN －γ可以显著升高 CXCL9和 CXCL10的表达水平，TNF －α、Poly（I∶C）、LPS 和 PHA 均可升高 CXCL9和／或 CXCL10的表达水平，与 IFN －γ有一定的协同作用，其中 Poly（I∶C）效果最强。IFN －γ、TNF －α、Poly（I∶C）、LPS 和 PHA 等刺激因子本身在单独或联合应用时对3种肿瘤细胞均无明显增殖抑制作用。相同刺激条件下，胃癌高中分化细胞株的 CXCL9和 CXCL10表达显著高于低分化细胞株。CIK、CAPRI 两种方式活化后细胞膜表面 CXCR3显著升高。CIK、CAPRI 两种活化方式刺激 CXCR3的升高效能相当，无统计学差异。3种血样[健康足月新生儿脐带血、健康成人外周血单个核细胞（PBMC）、肿瘤患者 PBMC]在相同活化方式刺激条件下 CXCR3的表达无统计学差异。免疫活性细胞经由 CXCL9、CX-CL10与 CXCR3介导，可以更多数量归巢至肿瘤。结论：趋化因子 CXCL9和 CXCL10及其受体 CXCR3的高表达可以有效促进免疫细胞靶向归巢至肿瘤。
목적：탐토불동자격인자대위암중 CXCL9화 CXCL10표체적영향，급불동혈양중면역세포경불동방식활화후，기막표면 CXCR3적표체정황，건립병평개 CXCR3화 CXCL9、CXCL10개도적파향항위암면역치료신기술。방법：채용 IFN －γ、TNF －α、Poly（I∶C）、LPS 화 PHA 자격위암세포（BGC －823、SGC －7901급 MKN－28），ELISA 법측정종류세포분비 CXCL9화 CXCL10적정황。장면역세포분별활화위 CIK 화 CAPRI，류식세포술검측활화전후면역세포표면 CXCR3적표체정황。사갑기우담서람（MTT）법평개자격인자[IFN －γ、TNF －α、Poly（I∶C）、LPS 화 PHA]본신대종류세포증식적영향。Transwell 추화실험비교추화인자 CXCL9화CXCL10급기수체 CXCR3적표체차이대면역세포파향귀소지종류적수량적영향。결과：위암세포주기출상태무명현 CXCL9화 CXCL10적표체，IFN －γ가이현저승고 CXCL9화 CXCL10적표체수평，TNF －α、Poly（I∶C）、LPS 화 PHA 균가승고 CXCL9화／혹 CXCL10적표체수평，여 IFN －γ유일정적협동작용，기중 Poly（I∶C）효과최강。IFN －γ、TNF －α、Poly（I∶C）、LPS 화 PHA 등자격인자본신재단독혹연합응용시대3충종류세포균무명현증식억제작용。상동자격조건하，위암고중분화세포주적 CXCL9화 CXCL10표체현저고우저분화세포주。CIK、CAPRI 량충방식활화후세포막표면 CXCR3현저승고。CIK、CAPRI 량충활화방식자격 CXCR3적승고효능상당，무통계학차이。3충혈양[건강족월신생인제대혈、건강성인외주혈단개핵세포（PBMC）、종류환자 PBMC]재상동활화방식자격조건하 CXCR3적표체무통계학차이。면역활성세포경유 CXCL9、CX-CL10여 CXCR3개도，가이경다수량귀소지종류。결론：추화인자 CXCL9화 CXCL10급기수체 CXCR3적고표체가이유효촉진면역세포파향귀소지종류。
Objective:The chemokine receptor CXCR3 and its corresponding ligands,including monokine induced by IFN -γ(CXCL9)and interferon -γinducible protein 10kDa (CXCL10),are known to impact immunocytes infil-tration.However,the optimal mode that regulates this infiltration requires further understanding.Here we investigated that how different regulators effect the expression level of CXCR3 or CXCL9 /10.Methods:For regulation studies, BGC -823,SGC -7901 and MKN -28 gastric cancer cells were stimulated with IFN -γ,TNF -α,Poly (I∶C),LPS and PHA.CXCL9 /CXCL10 released from cells was measured by ELISA.Flow cytometry was used to assess the ex-pression levels of CXCR3 on immunocytes before and after culture.The affinity of immunocytes to gastric cancer cells was determined by transwell migration assay.Results:CXCL9 and CXCL10 were prominently produced from 3 gastric cancer cell lines with impact of stimulants used respectively,except PHA,while they can not be detected in the culture medium of unstimulated cells.This effect was further enhanced by combination of IFN -γand the rest reagents.Mod-erately or well differentiated gastric cancer cells produced more chemokines than poorly differentiated cells with the same stimulants.There were higher expression levels of CXCR3 on immune cells after activation.And various sources of blood samples had difference of CXCR3 expression.Moreover,there was no significant difference between the two patterns(CIK and CAPRI group)in shaping immunocytes surface receptor portrait.Chemotaxis assay showed en-hanced transmigration of immunocytes to gastric cancer cells.Conclusion:Increase of the intratumoral CXCL9 /10 concentration and the surface express level of CXCR3 on immunocytes leads to higher affinity of immune cell to gastric cancer cells.CXCR3 and CXCL9 /CXCL10 might be the potential therapeutical candidates to strengthen the efficiency of gastric cancer immunotherapy.