中华病理学杂志
中華病理學雜誌
중화병이학잡지
Chinese Journal of Pathology
2012年
4期
248-253
,共6页
张璐%阿尔孜古丽·吐尔逊%古扎丽努尔·阿不力孜
張璐%阿爾孜古麗·吐爾遜%古扎麗努爾·阿不力孜
장로%아이자고려·토이손%고찰려노이·아불력자
宫颈肿瘤%肿瘤细胞,培养的%维吾尔族
宮頸腫瘤%腫瘤細胞,培養的%維吾爾族
궁경종류%종류세포,배양적%유오이족
Uterine cervical neoplasms%Tumor cells,cultured%Uygur nationality
目的 建立维吾尔族妇女宫颈癌细胞株并观察其生物学特性,为维吾尔族妇女宫颈癌的病因学及临床治疗途径研究提供实验材料.方法 无菌切除维吾尔族妇女低分化宫颈鳞状上皮细胞癌的手术标本,用酶消化法体外培养,连续传代稳定生长后,测定细胞活力,绘制细胞生长曲线.采用光镜、电镜观察细胞形态并进行细胞周期和染色体分析.观察Balb/c裸鼠接种后的成瘤情况.采用免疫荧光检测分子标志物CK17和CD44,兔超敏二步法检测分子标志物CK14、Ki-67和波形蛋白的表达情况.结果 酶消化法体外培养获得维吾尔族妇女宫颈癌细胞株,持续培养12个月,传代70代,细胞呈多边形,上皮样绌胞排列贴壁生长,无接触抑制,生长稳定,群体倍增时间为51.9h,瓶壁法和双层琼脂法的集落形成率分别为32.5％和15.6％.超微结构显示,细胞表面有较多突起或微绒毛,胞质内有大量杆状或棒状体和较典型的桥粒结构,核异型性明显.染色体数目在32 ～ 97条之间,众数在54～ 86条之间(60.3％),结构为人类染色体.裸鼠接种可形成移植瘤,组织病理形态学和患者原始肿瘤一致.细胞株分子标志物CK17、CD44、Ki-67、波形蛋白呈阳性表达,CK14呈阴性表达.结论 通过酶消化法体外培养建立的维吾尔族妇女宫颈癌细胞株具有人宫颈鳞状上皮细胞癌的特点,与原发癌保持相同的生物学特性,体外连续传代培养12个月,细胞形态及生长周期恒定,有望成为一个稳定的细胞株.
目的 建立維吾爾族婦女宮頸癌細胞株併觀察其生物學特性,為維吾爾族婦女宮頸癌的病因學及臨床治療途徑研究提供實驗材料.方法 無菌切除維吾爾族婦女低分化宮頸鱗狀上皮細胞癌的手術標本,用酶消化法體外培養,連續傳代穩定生長後,測定細胞活力,繪製細胞生長麯線.採用光鏡、電鏡觀察細胞形態併進行細胞週期和染色體分析.觀察Balb/c裸鼠接種後的成瘤情況.採用免疫熒光檢測分子標誌物CK17和CD44,兔超敏二步法檢測分子標誌物CK14、Ki-67和波形蛋白的錶達情況.結果 酶消化法體外培養穫得維吾爾族婦女宮頸癌細胞株,持續培養12箇月,傳代70代,細胞呈多邊形,上皮樣絀胞排列貼壁生長,無接觸抑製,生長穩定,群體倍增時間為51.9h,瓶壁法和雙層瓊脂法的集落形成率分彆為32.5％和15.6％.超微結構顯示,細胞錶麵有較多突起或微絨毛,胞質內有大量桿狀或棒狀體和較典型的橋粒結構,覈異型性明顯.染色體數目在32 ～ 97條之間,衆數在54～ 86條之間(60.3％),結構為人類染色體.裸鼠接種可形成移植瘤,組織病理形態學和患者原始腫瘤一緻.細胞株分子標誌物CK17、CD44、Ki-67、波形蛋白呈暘性錶達,CK14呈陰性錶達.結論 通過酶消化法體外培養建立的維吾爾族婦女宮頸癌細胞株具有人宮頸鱗狀上皮細胞癌的特點,與原髮癌保持相同的生物學特性,體外連續傳代培養12箇月,細胞形態及生長週期恆定,有望成為一箇穩定的細胞株.
목적 건립유오이족부녀궁경암세포주병관찰기생물학특성,위유오이족부녀궁경암적병인학급림상치료도경연구제공실험재료.방법 무균절제유오이족부녀저분화궁경린상상피세포암적수술표본,용매소화법체외배양,련속전대은정생장후,측정세포활력,회제세포생장곡선.채용광경、전경관찰세포형태병진행세포주기화염색체분석.관찰Balb/c라서접충후적성류정황.채용면역형광검측분자표지물CK17화CD44,토초민이보법검측분자표지물CK14、Ki-67화파형단백적표체정황.결과 매소화법체외배양획득유오이족부녀궁경암세포주,지속배양12개월,전대70대,세포정다변형,상피양출포배렬첩벽생장,무접촉억제,생장은정,군체배증시간위51.9h,병벽법화쌍층경지법적집락형성솔분별위32.5％화15.6％.초미결구현시,세포표면유교다돌기혹미융모,포질내유대량간상혹봉상체화교전형적교립결구,핵이형성명현.염색체수목재32 ～ 97조지간,음수재54～ 86조지간(60.3％),결구위인류염색체.라서접충가형성이식류,조직병리형태학화환자원시종류일치.세포주분자표지물CK17、CD44、Ki-67、파형단백정양성표체,CK14정음성표체.결론 통과매소화법체외배양건립적유오이족부녀궁경암세포주구유인궁경린상상피세포암적특점,여원발암보지상동적생물학특성,체외련속전대배양12개월,세포형태급생장주기항정,유망성위일개은정적세포주.
Objective To establish a uterine cervical carcinoma cell line of Uyghur ethnical background and to evaluate the related biological characteristics for future biomedical investigations of diseases in the Uyghur population.Methods Poorly-differentiated squamous cell carcinoma specimens of Uyghur patients were obtained and cultured in vitro by enzymatic digestion method,followed by continuous passaging to reach a stable growth determined by cell viability and growth curve.Morphological study,cell cycling and chromosomal analysis were performed.Tumorigenesis study was conducted by inoculation of nude mice.Biomarker( CK17,CD44,Ki-67,CK14 and vimentin)expression was detected by immunofluorescence and immunoeytoehemical tcchniqucs.Results A cervical carcinoma cell line was successfully established and maintained for 12 months through 70 passages. The cell line had a stable growth with a population doobling time of 51.9 h. Flask method and double agar-agar assay showed that the cell line had colonyforming rates of 32.5％ and 15.6％,respectively.Ultrastructural evaluation demonstrated numerous cell surface protrusions or microvilli,a large number of rod-shape structures in cytoplasm,typical dcsmosomes and nuclear atypia. Chromosomal analysis revealed human karyotype with the number of chromosomes per cell varying from 32-97 with a majority of 54-86 (60.3％ ).Xenogeneic tumors formed in nude mice showed histological structures identical to those of the primary tumor.The cells had high expression of CK17,CD44,Ki-67 and vimentin but no CK14 expression. Conclusions A cervical carcinoma cell line from a female Uyghur patient is successfully established.The cell line has the characteristics of human cervical squamous cell carcinoma,and it is stable with maintaining the characteristic biological and morphological features in vitro for more than 12 months,therefore,qualified as a stable cell line for further biomedical research.
