PDF(Pubmed)
Abstract：
UNASSIGNED: Intrahepatic cholangiocarcinoma (ICC) is an aggressive malignant tumor of the biliary tract that is prone to recurrence and metastasis and is characterized by poor sensitivity to chemotherapy and overall prognosis. For these reasons, there is an urgent need to understand its pathological mechanisms and develop effective treatments. To address this challenge, the establishment of suitable preclinical models is critical.
UNASSIGNED: Fresh ICC tissue samples were used for primary culture and subculture. The cell line was evaluated by cell proliferation assays, clonal formation assays, karyotype analysis, and short tandem repeat (STR) analysis. Drug resistances against oxaliplatin, paclitaxel, gemcitabine and 5-fluorouracil (5-FU) were evaluated by CCK-8 assay. Subcutaneous injection of 1 × 106 cells to three BALB/c nude mice was conducted for xenograft studies. The hematoxylin and eosin (H&E) staining was used to detect the pathological status of the cell line. The expression of biomarkers CK7, CK19, Ki-67, E-cadherin and vimentin was determined by immunocytochemistry assay.
UNASSIGNED: A new ICC cell line named ICC-X2 was successfully established. Like ICC-X3 established using the same patient\'s metastatic tumor, the cell line has been continuously cultured in vitro for more than a year and has been passaged more than 100 times. ICC-X2 retained the typical biliary epithelial morphology. The population doubling time of ICC-X2 is 48 h. The cells demonstrated an abnormal nearly tetraploid karyotype. The STR analysis confirmed that ICC-X2 was highly consistent with the primary tumor tissue and not cross-contaminated by existing cell lines. ICC-X2 cells positively expressed CK7, CK19, E-cadherin, and vimentin, and the positive expression of Ki-67 in ICC-X2 cells was 40%. The ICC-X2 cells exhibited a strong clonogenic ability. The drug sensitivity test indicated that ICC-X2 was sensitive to oxaliplatin and paclitaxel, but naturally resistant to gemcitabine and 5-FU. ICC-X2 was rapidly able to form transplanted tumors in vivo after subcutaneous inoculation in nude mice.
UNASSIGNED: ICC-X2 is an excellent experimental model that can be used for studying the occurrence, development, and metastasis of ICC and investigating the mechanism of tumor drug resistance.
UNASSIGNED: Fresh ICC tissue samples were used for primary culture and subculture. The cell line was evaluated by cell proliferation assays, clonal formation assays, karyotype analysis, and short tandem repeat (STR) analysis. Drug resistances against oxaliplatin, paclitaxel, gemcitabine and 5-fluorouracil (5-FU) were evaluated by CCK-8 assay. Subcutaneous injection of 1 × 106 cells to three BALB/c nude mice was conducted for xenograft studies. The hematoxylin and eosin (H&E) staining was used to detect the pathological status of the cell line. The expression of biomarkers CK7, CK19, Ki-67, E-cadherin and vimentin was determined by immunocytochemistry assay.
UNASSIGNED: A new ICC cell line named ICC-X2 was successfully established. Like ICC-X3 established using the same patient\'s metastatic tumor, the cell line has been continuously cultured in vitro for more than a year and has been passaged more than 100 times. ICC-X2 retained the typical biliary epithelial morphology. The population doubling time of ICC-X2 is 48 h. The cells demonstrated an abnormal nearly tetraploid karyotype. The STR analysis confirmed that ICC-X2 was highly consistent with the primary tumor tissue and not cross-contaminated by existing cell lines. ICC-X2 cells positively expressed CK7, CK19, E-cadherin, and vimentin, and the positive expression of Ki-67 in ICC-X2 cells was 40%. The ICC-X2 cells exhibited a strong clonogenic ability. The drug sensitivity test indicated that ICC-X2 was sensitive to oxaliplatin and paclitaxel, but naturally resistant to gemcitabine and 5-FU. ICC-X2 was rapidly able to form transplanted tumors in vivo after subcutaneous inoculation in nude mice.
UNASSIGNED: ICC-X2 is an excellent experimental model that can be used for studying the occurrence, development, and metastasis of ICC and investigating the mechanism of tumor drug resistance.
摘要：
■肝内胆管癌（ICC）是一种侵袭性的胆道恶性肿瘤，易于复发和转移，其特征是对化疗的敏感性和总体预后较差。由于这些原因,迫切需要了解其病理机制并开发有效的治疗方法。为了应对这一挑战，建立合适的临床前模型至关重要.
■新鲜的ICC组织样品用于原代培养和继代培养。通过细胞增殖试验评估细胞系，克隆形成试验,核型分析,和短串联重复(STR)分析。对奥沙利铂的耐药性，紫杉醇,通过CCK-8测定评估吉西他滨和5-氟尿嘧啶（5-FU）。对三只BALB/c裸鼠进行皮下注射1×106细胞用于异种移植研究。苏木精和伊红(H&E)染色用于检测细胞系的病理状况。免疫细胞化学法检测生物标志物CK7、CK19、Ki-67、E-cadherin和波形蛋白的表达。
■成功建立了名为ICC-X2的新ICC细胞系。就像使用同一患者的转移性肿瘤建立的ICC-X3一样，该细胞系已在体外连续培养超过一年，已传代超过100次。ICC-X2保留了典型的胆管上皮形态。ICC-X2的群体倍增时间为48小时。细胞表现出异常的近四倍体核型。STR分析证实，ICC-X2与原发性肿瘤组织高度一致，未被现有细胞系交叉污染。ICC-X2细胞阳性表达CK7，CK19，E-cadherin，还有波形蛋白,ICC-X2细胞中Ki-67的阳性表达率为40%。ICC-X2细胞表现出强的克隆形成能力。药敏试验提示ICC-X2对奥沙利铂和紫杉醇敏感,但对吉西他滨和5-FU具有天然抗性。ICC-X2在裸鼠皮下接种后能够在体内快速形成移植肿瘤。
■ICC-X2是一种出色的实验模型，可用于研究发生的情况，发展,和ICC的转移，探讨肿瘤耐药机制。
■新鲜的ICC组织样品用于原代培养和继代培养。通过细胞增殖试验评估细胞系，克隆形成试验,核型分析,和短串联重复(STR)分析。对奥沙利铂的耐药性，紫杉醇,通过CCK-8测定评估吉西他滨和5-氟尿嘧啶（5-FU）。对三只BALB/c裸鼠进行皮下注射1×106细胞用于异种移植研究。苏木精和伊红(H&E)染色用于检测细胞系的病理状况。免疫细胞化学法检测生物标志物CK7、CK19、Ki-67、E-cadherin和波形蛋白的表达。
■成功建立了名为ICC-X2的新ICC细胞系。就像使用同一患者的转移性肿瘤建立的ICC-X3一样，该细胞系已在体外连续培养超过一年，已传代超过100次。ICC-X2保留了典型的胆管上皮形态。ICC-X2的群体倍增时间为48小时。细胞表现出异常的近四倍体核型。STR分析证实，ICC-X2与原发性肿瘤组织高度一致，未被现有细胞系交叉污染。ICC-X2细胞阳性表达CK7，CK19，E-cadherin，还有波形蛋白,ICC-X2细胞中Ki-67的阳性表达率为40%。ICC-X2细胞表现出强的克隆形成能力。药敏试验提示ICC-X2对奥沙利铂和紫杉醇敏感,但对吉西他滨和5-FU具有天然抗性。ICC-X2在裸鼠皮下接种后能够在体内快速形成移植肿瘤。
■ICC-X2是一种出色的实验模型，可用于研究发生的情况，发展,和ICC的转移，探讨肿瘤耐药机制。
