中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2013年
6期
361-365
,共5页
司佩任%路筝%刘岩%马建霞%吴洪玉%李兆申
司珮任%路箏%劉巖%馬建霞%吳洪玉%李兆申
사패임%로쟁%류암%마건하%오홍옥%리조신
胰腺肿瘤%碘同位素%小剂量照射%肿瘤浸润
胰腺腫瘤%碘同位素%小劑量照射%腫瘤浸潤
이선종류%전동위소%소제량조사%종류침윤
Pancreatic neoplasms%Iodine isotopes%Low-level irradiation%Neoplasm invasiveness
目的 观察125I粒子短时低剂量率照射对胰腺癌Capan-2细胞神经浸润的影响,并探讨其分子机制.方法 建立胰腺癌Capan-2细胞和大鼠背根神经节(DRG)共培养及Capan-2或DRG单培养模型.通过125I粒子低剂量率照射平板对3种模型进行照射,以相应未照射模型作为对照.倒置显微镜下观察癌细胞、DRG的生长,图像分析软件计算神经突和癌细胞集落占据的表面积,ELISA法检测细胞培养上清液和基质胶溶解液中神经生长因子(NGF)及转化生长因子α(TGF-α)浓度,RT-PCR法检测胰腺癌Capan-2细胞神经营养因子-3(NT-3)mRNA表达.结果 共培养模型中DRG发出的神经突向癌细胞定向、集中生长,而癌细胞沿神经突的方向生长.经125I粒子照射后这种定向、集中和互逆的生长受到一定程度的抑制.共培养组第5天所增加的神经突表面积为290.15±12.08,较DRG单培养组的124.83±6.96显著增加(P＜0.01),经照射后的神经突表面积减少到201.53±12.20(P ＜0.01);所增加的Capan-2细胞表面积为300.47±12.99,较Capan-2细胞单培养组的199.30±8.60显著增加(P＜0.01),经照射后的Capan-2细胞表面积减少到202.35±7.97 (P ＜0.01).共培养组不表达NT-3mRNA,经照射后NT-3mRNA表达量为0.68±0.04(P ＜0.05).共培养组培养上清液中NGF及TGF-α浓度分别为(27.56 ±13.73)、(40.86±20.73) ng/ml,经照射后分别升高到(94.98±33.80)、(157.54±83.76) ng/ml,差异有统计学意义(P＜0.05或＜0.01).共培养组基质胶溶解液中NGF及TGF-α浓度分别为(60.42 ±33.03)、(64.39±21.52) ng/ml,经照射后分别升高到(132.52±53.01)、(138.38±83.58) ng/ml,其中NGF的差异有统计学意义(P＜0.05).结论 125I粒子短时低剂量率照射可以抑制胰腺癌和神经的交互作用,其机制可能与癌细胞促神经浸润介质NGF、TGF-α和NT-3等表达上调有关.
目的 觀察125I粒子短時低劑量率照射對胰腺癌Capan-2細胞神經浸潤的影響,併探討其分子機製.方法 建立胰腺癌Capan-2細胞和大鼠揹根神經節(DRG)共培養及Capan-2或DRG單培養模型.通過125I粒子低劑量率照射平闆對3種模型進行照射,以相應未照射模型作為對照.倒置顯微鏡下觀察癌細胞、DRG的生長,圖像分析軟件計算神經突和癌細胞集落佔據的錶麵積,ELISA法檢測細胞培養上清液和基質膠溶解液中神經生長因子(NGF)及轉化生長因子α(TGF-α)濃度,RT-PCR法檢測胰腺癌Capan-2細胞神經營養因子-3(NT-3)mRNA錶達.結果 共培養模型中DRG髮齣的神經突嚮癌細胞定嚮、集中生長,而癌細胞沿神經突的方嚮生長.經125I粒子照射後這種定嚮、集中和互逆的生長受到一定程度的抑製.共培養組第5天所增加的神經突錶麵積為290.15±12.08,較DRG單培養組的124.83±6.96顯著增加(P＜0.01),經照射後的神經突錶麵積減少到201.53±12.20(P ＜0.01);所增加的Capan-2細胞錶麵積為300.47±12.99,較Capan-2細胞單培養組的199.30±8.60顯著增加(P＜0.01),經照射後的Capan-2細胞錶麵積減少到202.35±7.97 (P ＜0.01).共培養組不錶達NT-3mRNA,經照射後NT-3mRNA錶達量為0.68±0.04(P ＜0.05).共培養組培養上清液中NGF及TGF-α濃度分彆為(27.56 ±13.73)、(40.86±20.73) ng/ml,經照射後分彆升高到(94.98±33.80)、(157.54±83.76) ng/ml,差異有統計學意義(P＜0.05或＜0.01).共培養組基質膠溶解液中NGF及TGF-α濃度分彆為(60.42 ±33.03)、(64.39±21.52) ng/ml,經照射後分彆升高到(132.52±53.01)、(138.38±83.58) ng/ml,其中NGF的差異有統計學意義(P＜0.05).結論 125I粒子短時低劑量率照射可以抑製胰腺癌和神經的交互作用,其機製可能與癌細胞促神經浸潤介質NGF、TGF-α和NT-3等錶達上調有關.
목적 관찰125I입자단시저제량솔조사대이선암Capan-2세포신경침윤적영향,병탐토기분자궤제.방법 건립이선암Capan-2세포화대서배근신경절(DRG)공배양급Capan-2혹DRG단배양모형.통과125I입자저제량솔조사평판대3충모형진행조사,이상응미조사모형작위대조.도치현미경하관찰암세포、DRG적생장,도상분석연건계산신경돌화암세포집락점거적표면적,ELISA법검측세포배양상청액화기질효용해액중신경생장인자(NGF)급전화생장인자α(TGF-α)농도,RT-PCR법검측이선암Capan-2세포신경영양인자-3(NT-3)mRNA표체.결과 공배양모형중DRG발출적신경돌향암세포정향、집중생장,이암세포연신경돌적방향생장.경125I입자조사후저충정향、집중화호역적생장수도일정정도적억제.공배양조제5천소증가적신경돌표면적위290.15±12.08,교DRG단배양조적124.83±6.96현저증가(P＜0.01),경조사후적신경돌표면적감소도201.53±12.20(P ＜0.01);소증가적Capan-2세포표면적위300.47±12.99,교Capan-2세포단배양조적199.30±8.60현저증가(P＜0.01),경조사후적Capan-2세포표면적감소도202.35±7.97 (P ＜0.01).공배양조불표체NT-3mRNA,경조사후NT-3mRNA표체량위0.68±0.04(P ＜0.05).공배양조배양상청액중NGF급TGF-α농도분별위(27.56 ±13.73)、(40.86±20.73) ng/ml,경조사후분별승고도(94.98±33.80)、(157.54±83.76) ng/ml,차이유통계학의의(P＜0.05혹＜0.01).공배양조기질효용해액중NGF급TGF-α농도분별위(60.42 ±33.03)、(64.39±21.52) ng/ml,경조사후분별승고도(132.52±53.01)、(138.38±83.58) ng/ml,기중NGF적차이유통계학의의(P＜0.05).결론 125I입자단시저제량솔조사가이억제이선암화신경적교호작용,기궤제가능여암세포촉신경침윤개질NGF、TGF-α화NT-3등표체상조유관.
Objective To investigate the effect of Iodine 125 seeds short time low dose rate irradiation on perineural invasion (PNI) in pancreatic cancer Capan-2 cells,and explore its molecular mechanism.Methods The co-culture model was established by co-culturing the dorsal root ganglion (DRG) of SD rat and Capzn-2 cells line,while Capan-2 culture model and DRG culture model was also established.Iodine 125 seeds short time low dose rate irradiation tablet was used for the 3 models,and the model without irradiation was used as control.Cancer cell and DRG growth was observed under inverted microscopy,surface of neurite and cell colony growth was determined by image analysis software.The concentration of nerve growth factor (NGF),transforming growth factor-α (TGF-α) in cell culture supernatant and matrigel solution was tested by ELISA,and the expression of neurotrophin-3 (NT-3) mRNA was detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR).Results In the co-culture model,neurite of DRG showed a direction to cancer cells and had a concentrated growth towards cancer cells.And Capan-2 cells formed more colonies towards neurite.However,in irradiation groups,the symbiotic phenomenon was inhibited to some degree.Increased surface of neurite in co-culture model at 5th day was 290.15 ± 12.08,which was significantly higher than that in DRG group (124.83 ± 6.96,P ＜ 0.01),but the surface of neurite was decreased to 201.53 ± 12.20 after irradiation (P ＜0.01).Increased surface of Capan-2 cell was 300.47 ± 12.99,which was significantly higher than that in Capan-2 group (199.30 ± 8.60,P ＜ 0.01),but the surface of Capan-2 was decreased to 202.35 ± 7.97 after irradiation (P ＜ 0.01).NT-3 mRNA was seldom or not expressed in supernatant of co-culture model,but it was strongly expressed (0.68 ± 0.04) after irradiation (P ＜ 0.05).The concentration of NGF and TGF-α in supernatant of co-culture model were (27.56 ± 13.73),(40.86 ± 20.73) ng/ml,after irradiation they were increased to (94.98 ± 33.80),(157.54 ± 83.76) ng,/ml,and the difference between the two groups was statistically significant (P＜0.05 or ＜0.01).The concentration of NGF and TGF-α in matrigel lysate of co-culture model were (60.42 ± 33.03),(64.39 ± 21.52)ng/ml,after irradiation they were increased to (132.52 ±53.01),(138.38 ±83.58)ng/ml,and the difference of NGF concentration between the two groups was statistically significant (P ＜ 0.05).Conclusions Iodine-125 seeds short-time low-dose rate irradiation could inhibit interactions between nerve and Capan-2 cells,and the mechanism may be related to up-regulation of cancer cells perineural invasion promoter NGF,TGF-α and NT-3.
