中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2008年
6期
456-459
,共4页
王庆伟%刘宏%梁业民%王涛%朱旭东
王慶偉%劉宏%樑業民%王濤%硃旭東
왕경위%류굉%량업민%왕도%주욱동
喉肿瘤%蛋白酶抑制药%放射疗法%NF-κB%细胞凋亡
喉腫瘤%蛋白酶抑製藥%放射療法%NF-κB%細胞凋亡
후종류%단백매억제약%방사요법%NF-κB%세포조망
Laryngeal neoplasms%Protease inhibitors%Radiotherapy%NF-kappa B%Apoptosis
目的 探讨蛋白酶抑制剂硼替佐米对喉鳞状细胞癌(简称鳞癌)Hep-2细胞的放射增敏作用及其机制.方法 体外培养的Hep-2细胞在100 nmol/L的硼替佐米作用2 h后,经过0、2、46、8及10 Gy照射后,检测其放射敏感性的变化,并通过建立的裸鼠移植瘤模型验证其体内放射增敏作用.利用Trans AMTM NF-κB P65活性测定试剂盒检测硼替佐米对Hep-2细胞辐射诱导NF-κB活化的影响;用流式细胞仪分析细胞周期及照射前后的细胞凋亡;Hoechst 33342荧光染色法观察细胞凋亡状况.结果 克隆形成实验显示硼替佐米可增加Hep-2细胞的放射敏感性34%.裸鼠移植瘤模型的肿瘤生长延迟放射增敏值为1.46.经2 Gy和8 Gy照射后2 h可诱导NF-κB活化,且存在剂量相关性(r=0.989,P<0.05).硼替佐米在0、2、8 Gy不同的照射剂量均能降低Hep-2细胞NF-κB的活性(t值分别为20.02、14.20、17.46,P<0.01);经硼替佐米干预的细胞有明显的G2-M期阻滞作用(t=22.31,P=0.000),并在照射后明显的细胞凋亡率增加(P值均<0.01).Hoecht 33342染色可见细胞核均匀染色,凋亡细胞的核凝集,呈强蓝色荧光,细胞核碎裂呈花瓣状.硼替佐米照射组凋亡比例高于单纯照射组.结论 硼替佐米通过可影响Hep-2细胞的细胞周期分布、诱导细胞凋亡及抑制NF-κB活化的途径,从而增加其对放射的敏感性.
目的 探討蛋白酶抑製劑硼替佐米對喉鱗狀細胞癌(簡稱鱗癌)Hep-2細胞的放射增敏作用及其機製.方法 體外培養的Hep-2細胞在100 nmol/L的硼替佐米作用2 h後,經過0、2、46、8及10 Gy照射後,檢測其放射敏感性的變化,併通過建立的裸鼠移植瘤模型驗證其體內放射增敏作用.利用Trans AMTM NF-κB P65活性測定試劑盒檢測硼替佐米對Hep-2細胞輻射誘導NF-κB活化的影響;用流式細胞儀分析細胞週期及照射前後的細胞凋亡;Hoechst 33342熒光染色法觀察細胞凋亡狀況.結果 剋隆形成實驗顯示硼替佐米可增加Hep-2細胞的放射敏感性34%.裸鼠移植瘤模型的腫瘤生長延遲放射增敏值為1.46.經2 Gy和8 Gy照射後2 h可誘導NF-κB活化,且存在劑量相關性(r=0.989,P<0.05).硼替佐米在0、2、8 Gy不同的照射劑量均能降低Hep-2細胞NF-κB的活性(t值分彆為20.02、14.20、17.46,P<0.01);經硼替佐米榦預的細胞有明顯的G2-M期阻滯作用(t=22.31,P=0.000),併在照射後明顯的細胞凋亡率增加(P值均<0.01).Hoecht 33342染色可見細胞覈均勻染色,凋亡細胞的覈凝集,呈彊藍色熒光,細胞覈碎裂呈花瓣狀.硼替佐米照射組凋亡比例高于單純照射組.結論 硼替佐米通過可影響Hep-2細胞的細胞週期分佈、誘導細胞凋亡及抑製NF-κB活化的途徑,從而增加其對放射的敏感性.
목적 탐토단백매억제제붕체좌미대후린상세포암(간칭린암)Hep-2세포적방사증민작용급기궤제.방법 체외배양적Hep-2세포재100 nmol/L적붕체좌미작용2 h후,경과0、2、46、8급10 Gy조사후,검측기방사민감성적변화,병통과건립적라서이식류모형험증기체내방사증민작용.이용Trans AMTM NF-κB P65활성측정시제합검측붕체좌미대Hep-2세포복사유도NF-κB활화적영향;용류식세포의분석세포주기급조사전후적세포조망;Hoechst 33342형광염색법관찰세포조망상황.결과 극륭형성실험현시붕체좌미가증가Hep-2세포적방사민감성34%.라서이식류모형적종류생장연지방사증민치위1.46.경2 Gy화8 Gy조사후2 h가유도NF-κB활화,차존재제량상관성(r=0.989,P<0.05).붕체좌미재0、2、8 Gy불동적조사제량균능강저Hep-2세포NF-κB적활성(t치분별위20.02、14.20、17.46,P<0.01);경붕체좌미간예적세포유명현적G2-M기조체작용(t=22.31,P=0.000),병재조사후명현적세포조망솔증가(P치균<0.01).Hoecht 33342염색가견세포핵균균염색,조망세포적핵응집,정강람색형광,세포핵쇄렬정화판상.붕체좌미조사조조망비례고우단순조사조.결론 붕체좌미통과가영향Hep-2세포적세포주기분포、유도세포조망급억제NF-κB활화적도경,종이증가기대방사적민감성.
Objective To determine if proteasome inhibitior bortezomib leads to enhanced radiation sensitivity of Hep-2 human laryngeal cancer cells and the relative mechanisms.Methods HeD-2 ceHs with or without bortezomib were irradiated at 0,2,4,6,8,and 10 Gy.Growth and clonogenic survival data were obtained to assess effects of treatment on radiosensitization.In vitro results were tested in vivo using a kit The distribution of cell cycle and apoptosis were evaluated by flow cytometry.Morphological evidence of apoptotic cells were observed with Hochest 33342.Results It decreased cell growth and clonogenie survival.A 34%increase in radiosensitivity was observed for cells treated with bortezomih and radiation. activation was induced by radiation and inhibited by pretreatment with bortezomib.and was in a dose-dependent manner(r=0.989.P<0.05).Hep-2 cells treated with 1130 nmol/L Bortezomib were arrested at G2.M phase(t=22.31,P=0.000)and resulted in a11 increased apoptosis with and without irradiation (P<0.01).Morphological evidence of apoptotic cells could be distinguished under the fluorescence microscope after staining with Hochest 33342.Many nuclear fragments were observed in Hep-2 cells with bortezomib.Conclusion Bortezomib could enhanced the radiosensitivity of Hep-2 laryngeal cancer cells by regulation of the distribution of cell cycle.
