CAJ | 학술논문

目的诱导并筛选具有放射耐受性的单克隆肝癌细胞亚株,为进一步研究细胞抗放射生物学变化构建实验模型.方法采用人肝癌HepG2细胞进行放射诱导,分次照射.累积吸收剂量为60 Gy.经细胞克隆筛选、建株.进行形态学和细胞超微结构观察,同时测定细胞生长特性以及放射敏感性变化与亲本HepG2细胞进行比较,并观察2 Gy照射后细胞内放射相关抗拒基因mRNA表达水平的变化,对该细胞亚株进行鉴定,定名为HepG2/R60细胞亚株.结果通过2 Gy×30次分割照射诱导,成功建立HepG2/R60细胞亚株.细胞鉴定结果显示,与亲本HepG2细胞比较,细胞形态不规则,伪足伸展,折光度清晰,细胞问连接较为松散.透射电镜观察HepG2/R60细胞表面微绒毛明显增多,线粒体丰富,高尔基体发达.细胞生长延缓,倍增时间明显延迟为34.9 h,与HepG2细胞比较,放射敏感性显著降低,放射相关抗拒基因表达明显升高.结论成功建立具有放射耐受性的人肝癌细胞亚株:HepG2/R60.经鉴定与其亲本的HepG2细胞比较,其放射敏感性显著降低.
목적 유도병사선구유방사내수성적단극륭간암세포아주,위진일보연구세포항방사생물학변화구건실험모형.방법 채용인간암HepG2세포진행방사유도,분차조사.루적흡수제량위60 Gy.경세포극륭사선、건주.진행형태학화세포초미결구관찰,동시측정세포생장특성이급방사민감성변화여친본HepG2세포진행비교,병관찰2 Gy조사후세포내방사상관항거기인mRNA표체수평적변화,대해세포아주진행감정,정명위HepG2/R60세포아주.결과 통과2 Gy×30차분할조사유도,성공건립HepG2/R60세포아주.세포감정결과현시,여친본HepG2세포비교,세포형태불규칙,위족신전,절광도청석,세포문련접교위송산.투사전경관찰HepG2/R60세포표면미융모명현증다,선립체봉부,고이기체발체.세포생장연완,배증시간명현연지위34.9 h,여HepG2세포비교,방사민감성현저강저,방사상관항거기인표체명현승고.결론 성공건립구유방사내수성적인간암세포아주:HepG2/R60.경감정여기친본적HepG2세포비교,기방사민감성현저강저.
Objective To induce and isolate the monoclonal cell subline, in order to establish the experimental model for further investigating biologic characteristics in radiotolerant hepatoma cells. Methods HepG2 cells were irradiated by γ-rays at the dose of 2 Gy each time with the total absorbed dose of 60 Gy. After monoclonal cell being selected and extensively cultured, the cell subline was named as HepG2/R60. Furthermore, HepG2/R60 cells were identified by observing the changes of morphology, ultrastructure, growth characteristics and radiosensitivity. The levels of radioresistant correlative gene mRNA in HepG2/R60 cells after exposure to 2 Gy irradiation, were also detected by RT-PCR, and then compared with parental HepG2 cells. Results HepG2/R60 cell subline was successfully established by fractionated irradiation at 2 Gy. HepG2/R60 cells displayed higher irregularity, the clearer appearance and dissociation of cell junctions compared with parental HepG2 cells. Ultrastnictural investigations through transmission electron microscopy (TEM) showed that there was an increase of microvillus on the surfaces of HepG2/R60 cells with plenty of rough endo-plasmic reticulum, abundance of mitochondria and viable Golgi complex. Further observation found that the growth of HepG2/R60 cells was slower and its population doubling time (PDT) prolonged arrived at 34.9 h. Moreover, the radiosensitivity of HepG2/R60 cells was lower than that of parental HepG2 cells. Additionally, the levels of radioresistance correlative genes were increased in HepG2/R60 cells by 2 Gy irradiaiton Conclusions Radiotolerant cell subline - HepG2/R60 was successfully isolated and established by fractionated irradiation.