中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2014年
6期
435-439
,共5页
陈应泰%王成锋%何明彦%张红%陈学忠
陳應泰%王成鋒%何明彥%張紅%陳學忠
진응태%왕성봉%하명언%장홍%진학충
胰腺肿瘤%外周血%低剂量重离子束照射%T细胞%NK细胞%免疫
胰腺腫瘤%外週血%低劑量重離子束照射%T細胞%NK細胞%免疫
이선종류%외주혈%저제량중리자속조사%T세포%NK세포%면역
Pancreatic neoplasms%Peripheral blood%Low dose heavy ion irradiation%T cells%NK cells%Immunity
目的 研究低剂量重离子束(12C6+)照射对胰腺癌患者外周血淋巴细胞(PBL)亚群比例和细胞因子表达的影响.方法 采集21例胰腺癌患者的PBL,每例患者标本分为假照组、X线照射组(0.05 Gy)和12C6+照射组(0.05 Gy)进行照射.照射后24 h,采用流式细胞仪检测T细胞与NK细胞亚群比例变化,采用实时定量PCR法检测白介素2(IL-2)、γ干扰素(IFN-γ)和肿瘤坏死因子α(TNF-α) mRNA的表达,采用酶联免疫吸附试验检测IL-2、IFN-γY和TNF-α蛋白的表达水平.结果 12C6+照射组外周血中CD3+T细胞、CD3+ CD4+T细胞和CD3+ CD8+T细胞的比例分别为(67.15±4.36)％、(19.02±2.35)％和(46.59±6.07)％,X线照射组外周血中CD3+T细胞、CD3+ CD4+T细胞和CD3+ CD8+T细胞的比例分别为(60.81±8.35)％、(17.21±2.86)％和(41.18±6.35)％,均明显高于假照组(均P ＜0.05).但假照组、X线照射组和12C6+照射组CD3+ CD4+T细胞与CD3+ CD8+T细胞的比值分别为0.67、0.65和0.68,差异无统计学意义(P＞0.05).与假照组比较,X线照射组和12C6+照射组外周血中CD3-CD16+ NK细胞、CD3-CD56+ NK细胞和CD16+ CD56+ NK细胞的比例均未发生明显变化(均P＞0.05).12C6+照射组外周血淋巴细胞中IFN-γ和IL-2 mRNA表达量的Ct值分别为23.35±3.16和24.19±3.56,X线照射组外周血淋巴细胞中IFN-γ和IL-2 mRNA表达量的Ct值分别为27.25±2.15和27.85±4.08,与假照组的差异均有统计学意义(均P＜0.05);但TNF-αmRNA在各组间的表达差异无统计学意义(P＞0.05).假照组、X线照射组和12C6+照射组外周血淋巴细胞中IFN-γ、IL-2和TNF-α蛋白的表达与mRNA的表达趋势一致.结论 低剂量重离子束照射能明显减轻胰腺癌患者的免疫抑制,且较传统的X线照射有更大的优势.T细胞亚群和细胞因子的表达水平可作为低剂量辐射效应的评价指标.
目的 研究低劑量重離子束(12C6+)照射對胰腺癌患者外週血淋巴細胞(PBL)亞群比例和細胞因子錶達的影響.方法 採集21例胰腺癌患者的PBL,每例患者標本分為假照組、X線照射組(0.05 Gy)和12C6+照射組(0.05 Gy)進行照射.照射後24 h,採用流式細胞儀檢測T細胞與NK細胞亞群比例變化,採用實時定量PCR法檢測白介素2(IL-2)、γ榦擾素(IFN-γ)和腫瘤壞死因子α(TNF-α) mRNA的錶達,採用酶聯免疫吸附試驗檢測IL-2、IFN-γY和TNF-α蛋白的錶達水平.結果 12C6+照射組外週血中CD3+T細胞、CD3+ CD4+T細胞和CD3+ CD8+T細胞的比例分彆為(67.15±4.36)％、(19.02±2.35)％和(46.59±6.07)％,X線照射組外週血中CD3+T細胞、CD3+ CD4+T細胞和CD3+ CD8+T細胞的比例分彆為(60.81±8.35)％、(17.21±2.86)％和(41.18±6.35)％,均明顯高于假照組(均P ＜0.05).但假照組、X線照射組和12C6+照射組CD3+ CD4+T細胞與CD3+ CD8+T細胞的比值分彆為0.67、0.65和0.68,差異無統計學意義(P＞0.05).與假照組比較,X線照射組和12C6+照射組外週血中CD3-CD16+ NK細胞、CD3-CD56+ NK細胞和CD16+ CD56+ NK細胞的比例均未髮生明顯變化(均P＞0.05).12C6+照射組外週血淋巴細胞中IFN-γ和IL-2 mRNA錶達量的Ct值分彆為23.35±3.16和24.19±3.56,X線照射組外週血淋巴細胞中IFN-γ和IL-2 mRNA錶達量的Ct值分彆為27.25±2.15和27.85±4.08,與假照組的差異均有統計學意義(均P＜0.05);但TNF-αmRNA在各組間的錶達差異無統計學意義(P＞0.05).假照組、X線照射組和12C6+照射組外週血淋巴細胞中IFN-γ、IL-2和TNF-α蛋白的錶達與mRNA的錶達趨勢一緻.結論 低劑量重離子束照射能明顯減輕胰腺癌患者的免疫抑製,且較傳統的X線照射有更大的優勢.T細胞亞群和細胞因子的錶達水平可作為低劑量輻射效應的評價指標.
목적 연구저제량중리자속(12C6+)조사대이선암환자외주혈림파세포(PBL)아군비례화세포인자표체적영향.방법 채집21례이선암환자적PBL,매례환자표본분위가조조、X선조사조(0.05 Gy)화12C6+조사조(0.05 Gy)진행조사.조사후24 h,채용류식세포의검측T세포여NK세포아군비례변화,채용실시정량PCR법검측백개소2(IL-2)、γ간우소(IFN-γ)화종류배사인자α(TNF-α) mRNA적표체,채용매련면역흡부시험검측IL-2、IFN-γY화TNF-α단백적표체수평.결과 12C6+조사조외주혈중CD3+T세포、CD3+ CD4+T세포화CD3+ CD8+T세포적비례분별위(67.15±4.36)％、(19.02±2.35)％화(46.59±6.07)％,X선조사조외주혈중CD3+T세포、CD3+ CD4+T세포화CD3+ CD8+T세포적비례분별위(60.81±8.35)％、(17.21±2.86)％화(41.18±6.35)％,균명현고우가조조(균P ＜0.05).단가조조、X선조사조화12C6+조사조CD3+ CD4+T세포여CD3+ CD8+T세포적비치분별위0.67、0.65화0.68,차이무통계학의의(P＞0.05).여가조조비교,X선조사조화12C6+조사조외주혈중CD3-CD16+ NK세포、CD3-CD56+ NK세포화CD16+ CD56+ NK세포적비례균미발생명현변화(균P＞0.05).12C6+조사조외주혈림파세포중IFN-γ화IL-2 mRNA표체량적Ct치분별위23.35±3.16화24.19±3.56,X선조사조외주혈림파세포중IFN-γ화IL-2 mRNA표체량적Ct치분별위27.25±2.15화27.85±4.08,여가조조적차이균유통계학의의(균P＜0.05);단TNF-αmRNA재각조간적표체차이무통계학의의(P＞0.05).가조조、X선조사조화12C6+조사조외주혈림파세포중IFN-γ、IL-2화TNF-α단백적표체여mRNA적표체추세일치.결론 저제량중리자속조사능명현감경이선암환자적면역억제,차교전통적X선조사유경대적우세.T세포아군화세포인자적표체수평가작위저제량복사효응적평개지표.
Objective The aim of this study was to examine the effect of low dose heavy ion irradiation on the subset percentage and expression of cytokines of peripheral blood lymphocytes (PBL) in patients with pancreatic cancer.Methods PBL from 21 patients with pancreatic cancer were divided into three groups:sham,X-ray and 12C6+ irradiation groups,and the cell responses were measured at 24 hours after radiation exposure.The percentages of T and NK cell subsets were detected by flow cytometry.The mRNA expression of interleukin (IL)-2,tumor necrosis factor (TNF)-α and interferon (IFN)-γ were examined by real-time quantitative RT-PCR (qRT-PCR).The cytokine protein levels in supernatant of cultured cells were assayed by enzyme-linked immunosorbent assays (ELISA).Results The percentage of T lymphocyte subsets was significantly increased at 24 hours after exposure to low dose radiation,and the effect was more pronounced in the group receiving 0.05 Gy 12C6+ ion irradiation than that in the group receiving X-ray irradiation [CD3 + T cells:(67.15 ± 4.36) ％ vs.(60.81 ± 8.35) ％ ; CD3 + CD4 + T cells:(19.02±2.35)％ vs.(17.21±2.86)％; CD3+CD8+ T cells:(46.59±6.07)％ vs.(41.18 ± 6.35) ％.(P ＜ 0.05 for all)].However,there were no significant changes in the CD3 + CD4 +/CD3 + CD8 + ratio (0.67 for sham,0.65 for X-ray,and 0.68 for 12C6+ groups) and percentage of NK cell subsets (P ＞0.05 for all).Expression levels of IFN-γ mRNA (cycle threshold/CT value was 23.35 ± 3.16 for 12C6+,CT value was 27.25 ± 2.15 for X-ray) and IL-2 (CT value was 24.19 ± 3.56 for 12C6+,CT value was 27.85 ±4.08 for X-ray) in PBL,and their protein levels in the supernatant were significantly increased at 24 hours after exposure to the low dose radiation (P ＜ 0.05).The effects were more pronounced in the group receiving 0.05 Gy 12C6+ ion irradiation than that in the group receiving X-ray irradiation.However,there was no significant change in the TNF-α production of PBL.Conclusions Low dose irradiation may alleviate the immune suppression caused by tumor burden and that the effect is more pronounced for 0.05 Gy high linear energy transfer (LET) 12C6+ irradiation.The percentage of T cell subsets and cytokines production could be used as sensitive indicators of acute response to low dose irradiation.
