中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2015年
38期
3142-3146
,共5页
姚懿雯%吴军录%权文强%万海英%李冬
姚懿雯%吳軍錄%權文彊%萬海英%李鼕
요의문%오군록%권문강%만해영%리동
肺癌%抗菌肽%炎症%巨噬细胞
肺癌%抗菌肽%炎癥%巨噬細胞
폐암%항균태%염증%거서세포
Lung cancer%Antimicrobial peptide%Inflammation%Macrophage
目的 探讨非肿瘤细胞分泌的抗菌肽cathelicidin促进肺癌生长的作用及其机制.方法 Lewis肺癌细胞(LLCl)通过尾静脉注射入cathelicidin敲除小鼠(CRAMP-/-)和对照小鼠(WT)建立小鼠转移性肺癌模型.对小鼠肺重量及表面肿瘤个数进行评价.利用Kaplan-Meier(K-M)生存率曲线分析小鼠生存率.免疫组织化学方法检测小鼠cathelicidin、肿瘤增殖抗原Ki-67和CD68表达;涂片染色计数肺泡灌洗液中各种炎症细胞个数.结果 Cathlicidin在肿瘤组织的炎症免疫细胞中呈高表达状态,而在肿瘤细胞中表达很弱.CRAMP-/-小鼠的肺重量和肿瘤数分别为(0.25±0.04)g和(9.60±2.25)个,均显著低于WT小鼠的(0.65±0.05)g和(23.40±2.68)个(t=6.07、3.95,均P＜0.05).K-M生存率分析结果显示,CRAMP-/-小鼠的中位生存时间为49(46 ～51)d,长于Wr小鼠的34(28 ～39) d(x2 =12.00,P＜0.05),且组织切片中Ki-67阳性肿瘤细胞为(18.80±2.38)％,明显低于WT小鼠的(35.80±2.96)％(=4.48,P＜0.05).肺泡灌洗液炎症细胞计数显示,CRAMP-/-小鼠的总细胞、中性粒细胞、淋巴细胞及巨噬细胞数量分别为(4.72±0.86)×104个、(0.08±0.02)×104个、(0.05±0.02) ×104个和(4.60±0.84)×104个,均低于WT小鼠的(16.18±1.61)×104个、(0.32±0.05)×104个、(0.20±0.05)×104个和(15.66±1.57)×104个(t=6.28、4.39、3.00、6.20,均P＜0.05),其中以巨噬细胞数量下降最为明显;免疫组织化学检测结果显示,在肿瘤组织中CRAMP-/-小鼠巨噬细胞的数量为(6.77±3.12)个/高倍视野,明显低于WT小鼠的(15.53±2.28)个/高倍视野(t=3.41,P＜0.05).结论 非肿瘤细胞分泌的cathelicidin具有促进小鼠肿瘤细胞增殖及肿瘤生长的作用,募集炎症细胞如巨噬细胞进入肿瘤微环境可能是其主要作用机制.
目的 探討非腫瘤細胞分泌的抗菌肽cathelicidin促進肺癌生長的作用及其機製.方法 Lewis肺癌細胞(LLCl)通過尾靜脈註射入cathelicidin敲除小鼠(CRAMP-/-)和對照小鼠(WT)建立小鼠轉移性肺癌模型.對小鼠肺重量及錶麵腫瘤箇數進行評價.利用Kaplan-Meier(K-M)生存率麯線分析小鼠生存率.免疫組織化學方法檢測小鼠cathelicidin、腫瘤增殖抗原Ki-67和CD68錶達;塗片染色計數肺泡灌洗液中各種炎癥細胞箇數.結果 Cathlicidin在腫瘤組織的炎癥免疫細胞中呈高錶達狀態,而在腫瘤細胞中錶達很弱.CRAMP-/-小鼠的肺重量和腫瘤數分彆為(0.25±0.04)g和(9.60±2.25)箇,均顯著低于WT小鼠的(0.65±0.05)g和(23.40±2.68)箇(t=6.07、3.95,均P＜0.05).K-M生存率分析結果顯示,CRAMP-/-小鼠的中位生存時間為49(46 ～51)d,長于Wr小鼠的34(28 ～39) d(x2 =12.00,P＜0.05),且組織切片中Ki-67暘性腫瘤細胞為(18.80±2.38)％,明顯低于WT小鼠的(35.80±2.96)％(=4.48,P＜0.05).肺泡灌洗液炎癥細胞計數顯示,CRAMP-/-小鼠的總細胞、中性粒細胞、淋巴細胞及巨噬細胞數量分彆為(4.72±0.86)×104箇、(0.08±0.02)×104箇、(0.05±0.02) ×104箇和(4.60±0.84)×104箇,均低于WT小鼠的(16.18±1.61)×104箇、(0.32±0.05)×104箇、(0.20±0.05)×104箇和(15.66±1.57)×104箇(t=6.28、4.39、3.00、6.20,均P＜0.05),其中以巨噬細胞數量下降最為明顯;免疫組織化學檢測結果顯示,在腫瘤組織中CRAMP-/-小鼠巨噬細胞的數量為(6.77±3.12)箇/高倍視野,明顯低于WT小鼠的(15.53±2.28)箇/高倍視野(t=3.41,P＜0.05).結論 非腫瘤細胞分泌的cathelicidin具有促進小鼠腫瘤細胞增殖及腫瘤生長的作用,募集炎癥細胞如巨噬細胞進入腫瘤微環境可能是其主要作用機製.
목적 탐토비종류세포분비적항균태cathelicidin촉진폐암생장적작용급기궤제.방법 Lewis폐암세포(LLCl)통과미정맥주사입cathelicidin고제소서(CRAMP-/-)화대조소서(WT)건립소서전이성폐암모형.대소서폐중량급표면종류개수진행평개.이용Kaplan-Meier(K-M)생존솔곡선분석소서생존솔.면역조직화학방법검측소서cathelicidin、종류증식항원Ki-67화CD68표체;도편염색계수폐포관세액중각충염증세포개수.결과 Cathlicidin재종류조직적염증면역세포중정고표체상태,이재종류세포중표체흔약.CRAMP-/-소서적폐중량화종류수분별위(0.25±0.04)g화(9.60±2.25)개,균현저저우WT소서적(0.65±0.05)g화(23.40±2.68)개(t=6.07、3.95,균P＜0.05).K-M생존솔분석결과현시,CRAMP-/-소서적중위생존시간위49(46 ～51)d,장우Wr소서적34(28 ～39) d(x2 =12.00,P＜0.05),차조직절편중Ki-67양성종류세포위(18.80±2.38)％,명현저우WT소서적(35.80±2.96)％(=4.48,P＜0.05).폐포관세액염증세포계수현시,CRAMP-/-소서적총세포、중성립세포、림파세포급거서세포수량분별위(4.72±0.86)×104개、(0.08±0.02)×104개、(0.05±0.02) ×104개화(4.60±0.84)×104개,균저우WT소서적(16.18±1.61)×104개、(0.32±0.05)×104개、(0.20±0.05)×104개화(15.66±1.57)×104개(t=6.28、4.39、3.00、6.20,균P＜0.05),기중이거서세포수량하강최위명현;면역조직화학검측결과현시,재종류조직중CRAMP-/-소서거서세포적수량위(6.77±3.12)개/고배시야,명현저우WT소서적(15.53±2.28)개/고배시야(t=3.41,P＜0.05).결론 비종류세포분비적cathelicidin구유촉진소서종류세포증식급종류생장적작용,모집염증세포여거서세포진입종류미배경가능시기주요작용궤제.
Objective To investigate the effect of antimicrobial peptide cathelicidin secreted by non-tumorous cells in lung tumor growth.Methods CRAMP-/-mice and WT mice were used to establish a lung cancer model via tail vein injection of Lewis lung carcinoma cells (LLC1).Lung was weighted and tumor number on the lung surface was counted.Kaplan-Meier (K-M) survival curve was used to analyze survival rate of mice.Expression of cathelicidin,Ki-67 and CD68 in the tumor tissue was measured by immunohistochemical analysis.BALF cells were stained with Diff Quik and percentages of leukocyte types were determined by light microscopy.Results Cathelicidin was high expression in inflammatory cells of tumor tissue,whereas weak expression in tumor cells.The lung weight and number of tumor in CRAMP-/-mice were (0.25 ± 0.04) g and (9.60 ± 2.25),respectively,which were significantly lower than those of WT mice (0.65 ± 0.05) g and (23.40 ± 2.68).The difference was statistically significant (t =6.07,3.95,all P ＜0.05).And Kaplan-Meier survival analysis showed median survival time of CRAMP-/-mice was 49 (46-51) d,which was longer than 34 (28-39) d of WT mice (x2 =12.00,P ＜ 0.05).And the positive rate of Ki-67 tumor cells was significant reduced from (35.80 ± 2.96) ％ in WT mice to (18.80 ± 2.38) ％ in CRAMP-/-groups (t =4.48,P ＜ 0.05).The total cell number as well as the number of lymphocytes,neutrophils,and macrophages in BALFs of CRAMP-/-mice were (4.72± 0.86) × 104,(0.08-0.02) × 104,(0.05 ± 0.02) × 104 and (4.60 ± 0.84) × 104,respectively,while of WT mice were (16.18±1.61) ×l04,(0.32±0.05) ×l04,(0.20±0.05) ×104 and (15.66±1.57) ×l04.All of them had significant difference(t =6.28,4.39,3.00,6.20,all P ＜0.05).In addition,the infiltration of macrophages into lung tumors was decreased in CRAMP-/-mice compared to WT mice,from (15.53 ± 2.28)/high power field to (6.77 ±3.12)/high power field (t =3.41,P ＜0.05).Conclusions Nontumor cells secreted cathelicidin promotes tumor cell proliferation and lung tumor growth.Recruitment of inflammatory cells such as macrophages into the tumor microenvironment may be the main mechanism of action.
