中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2012年
6期
583-587
,共5页
电离辐射%Toll样受体4%肿瘤细胞%放射敏感性
電離輻射%Toll樣受體4%腫瘤細胞%放射敏感性
전리복사%Toll양수체4%종류세포%방사민감성
Ionizing radiation%Toll-like receptor-4%Tumor cells%Radiosensitivity
目的 观察Toll样受体4(TLR4)对肿瘤细胞放射敏感性的影响.方法 将体外培养的RAW264.7、Lewis、MFC、Hepal-6、B16和NIH3T3细胞各分为假照组和5 Gy照射组,采用流式细胞术检测照射24 h后TLR4表达变化,从中筛选出照射后高表达和低表达TLR4的细胞,将其各自分为TAK242阻滞组、LPS刺激组和空白对照组,采用CCK-8试剂盒检测其增殖活性,用克隆形成实验计算其细胞存活分数,用Annexin V凋亡试剂盒检测其凋亡率,用流式细胞术观察细胞周期进程的变化.结果 5 Gy照射24 h后Lewis细胞TLR4表达水平明显高于照射前(t=-8.68,P＜0.01),MFC细胞则相反,照射后TLR4表达水平明显低于照射前(t=25.80,P＜0.01),而RAW264.7、Hepa1-6、B16细胞的TLR4表达水平比照射前有升高趋势,但无明显差异.5 Gy照射后Lewis和MFC细胞的增殖活性均明显降低(t=57.62、-6.23,P＜0.01),而用TAK242阻滞TLR4后Leiws细胞增殖活性更加降低(t=5.96,P＜0.01),但MFC细胞增殖活性反而明显升高(t=4.16,P＜0.01).5 Gy照射后Lewis和MFC细胞的存活分数均明显降低(t=13.37、19.24,P＜0.01),用TAK242阻滞TLR4后Leiws和MFC细胞存活分数更加降低(t=4.90、4.42,P＜0.05).5 Gy照射后Lewis细胞的凋亡率明显高于假照组(t=-167.85,P＜0.01),阻断TLR4后明显高于单纯照射组(t=-4.73,P＜0.01).照射后MFC细胞的凋亡率升高(t=-26.45,P＜0.01),阻断和激动TLR4其凋亡率均显著低于单纯照射组(t=8.87、-3.05,P＜0.05).Lewis和MFC细胞受照后G0/G1期、S期细胞明显降低(t=8.68、14.80、20.31、4.48,P＜0.01),G2/M期细胞上升(t=-37.48、-13.06,P＜0.01).两种细胞的TAK242阻断组和LPS刺激组各周期进程均无明显变化.结论 Lewis细胞的TLR4高表达与其受照后的增殖活性和凋亡率变化有关,而与其周期进程无关,TLR4影响肿瘤细胞的放射敏感性.
目的 觀察Toll樣受體4(TLR4)對腫瘤細胞放射敏感性的影響.方法 將體外培養的RAW264.7、Lewis、MFC、Hepal-6、B16和NIH3T3細胞各分為假照組和5 Gy照射組,採用流式細胞術檢測照射24 h後TLR4錶達變化,從中篩選齣照射後高錶達和低錶達TLR4的細胞,將其各自分為TAK242阻滯組、LPS刺激組和空白對照組,採用CCK-8試劑盒檢測其增殖活性,用剋隆形成實驗計算其細胞存活分數,用Annexin V凋亡試劑盒檢測其凋亡率,用流式細胞術觀察細胞週期進程的變化.結果 5 Gy照射24 h後Lewis細胞TLR4錶達水平明顯高于照射前(t=-8.68,P＜0.01),MFC細胞則相反,照射後TLR4錶達水平明顯低于照射前(t=25.80,P＜0.01),而RAW264.7、Hepa1-6、B16細胞的TLR4錶達水平比照射前有升高趨勢,但無明顯差異.5 Gy照射後Lewis和MFC細胞的增殖活性均明顯降低(t=57.62、-6.23,P＜0.01),而用TAK242阻滯TLR4後Leiws細胞增殖活性更加降低(t=5.96,P＜0.01),但MFC細胞增殖活性反而明顯升高(t=4.16,P＜0.01).5 Gy照射後Lewis和MFC細胞的存活分數均明顯降低(t=13.37、19.24,P＜0.01),用TAK242阻滯TLR4後Leiws和MFC細胞存活分數更加降低(t=4.90、4.42,P＜0.05).5 Gy照射後Lewis細胞的凋亡率明顯高于假照組(t=-167.85,P＜0.01),阻斷TLR4後明顯高于單純照射組(t=-4.73,P＜0.01).照射後MFC細胞的凋亡率升高(t=-26.45,P＜0.01),阻斷和激動TLR4其凋亡率均顯著低于單純照射組(t=8.87、-3.05,P＜0.05).Lewis和MFC細胞受照後G0/G1期、S期細胞明顯降低(t=8.68、14.80、20.31、4.48,P＜0.01),G2/M期細胞上升(t=-37.48、-13.06,P＜0.01).兩種細胞的TAK242阻斷組和LPS刺激組各週期進程均無明顯變化.結論 Lewis細胞的TLR4高錶達與其受照後的增殖活性和凋亡率變化有關,而與其週期進程無關,TLR4影響腫瘤細胞的放射敏感性.
목적 관찰Toll양수체4(TLR4)대종류세포방사민감성적영향.방법 장체외배양적RAW264.7、Lewis、MFC、Hepal-6、B16화NIH3T3세포각분위가조조화5 Gy조사조,채용류식세포술검측조사24 h후TLR4표체변화,종중사선출조사후고표체화저표체TLR4적세포,장기각자분위TAK242조체조、LPS자격조화공백대조조,채용CCK-8시제합검측기증식활성,용극륭형성실험계산기세포존활분수,용Annexin V조망시제합검측기조망솔,용류식세포술관찰세포주기진정적변화.결과 5 Gy조사24 h후Lewis세포TLR4표체수평명현고우조사전(t=-8.68,P＜0.01),MFC세포칙상반,조사후TLR4표체수평명현저우조사전(t=25.80,P＜0.01),이RAW264.7、Hepa1-6、B16세포적TLR4표체수평비조사전유승고추세,단무명현차이.5 Gy조사후Lewis화MFC세포적증식활성균명현강저(t=57.62、-6.23,P＜0.01),이용TAK242조체TLR4후Leiws세포증식활성경가강저(t=5.96,P＜0.01),단MFC세포증식활성반이명현승고(t=4.16,P＜0.01).5 Gy조사후Lewis화MFC세포적존활분수균명현강저(t=13.37、19.24,P＜0.01),용TAK242조체TLR4후Leiws화MFC세포존활분수경가강저(t=4.90、4.42,P＜0.05).5 Gy조사후Lewis세포적조망솔명현고우가조조(t=-167.85,P＜0.01),조단TLR4후명현고우단순조사조(t=-4.73,P＜0.01).조사후MFC세포적조망솔승고(t=-26.45,P＜0.01),조단화격동TLR4기조망솔균현저저우단순조사조(t=8.87、-3.05,P＜0.05).Lewis화MFC세포수조후G0/G1기、S기세포명현강저(t=8.68、14.80、20.31、4.48,P＜0.01),G2/M기세포상승(t=-37.48、-13.06,P＜0.01).량충세포적TAK242조단조화LPS자격조각주기진정균무명현변화.결론 Lewis세포적TLR4고표체여기수조후적증식활성화조망솔변화유관,이여기주기진정무관,TLR4영향종류세포적방사민감성.
Objective To investigate the effects of TLR4 on the radiosensitivity of tumor cells.Methods The cell lines of RAW264.7,Lewis,MFC,Hepal-6,Bl6,and NIH3T3 were irradiated with 5 Gy X-rays or sham-irradiated.24 h after irradiation,the expression of TLR4 was detected by flow cytometry.According to the TKR4 level,cells were divided into three groups:without treatment,LPS stimulation and TAK242 block.CCK-8 kit and Annexin-V Apoptosis Kit were used to detect cell proliferation,apoptosis and cell cycle distribution of each group.Results After 24 h of 5 Gy ionizing radiation,TLR4 was significantly increased in Lewis cells (t =-8.68,P ＜0.01) but decreased in MFC cells (t =25.8,P ＜ 0.01) and had no significant changes in Hepal-6,B16 and RAW264.7 cells.In addition,the proliferation vitality (t =57.62,-6.23,P ＜ 0.01) and survival fraction (t =13.37,19.24,P ＜ 0.01) of the Lewis and MFC cells were reduced especially for the TLR4-blocked cells,and the apoptosis rates of both Lewis (t=-167.85,P＜0.01) and MFC cells (t=-26.45,P＜0.01) were elevated.The percentages of G0/G1 phase and S phase Lewis cells were significant increased (t =8.68,14.89,P ＜ 0.01) but its G2/M phase were reduced (t =-37.48,P ＜ 0.01).However,the percentages of G0/G1 phase and S phase MFC cells were obviously reduced (t =20.31,4.48,P ＜ 0.01) and G2/M phase increased (t =-13.06,P ＜ 0.01).For both cell lines of Lewis and MFC,the cycle distribution of TAK242 and LPS groups didn't change significantly.Conclusions High expression TLR4 in the Lewis cells is related to cell proliferation and apoptosis but not cell cycle distribution,and hence TLR4 could influence the radiosensitivity of tumor cells.
