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Abstract：
BACKGROUND: Extrahepatic cholangiocarcinoma sarcoma is extremely rare in clinical practice. These cells consist of both epithelial and mesenchymal cells. Patient-derived cell lines that maintain tumor characteristics are valuable tools for studying the molecular mechanisms associated with carcinosarcoma. However, cholangiocarcinoma sarcoma cell lines are not available in cell banks.
OBJECTIVE: To establish and characterize a new extrahepatic cholangiocarcinoma sarcoma cell line, namely CBC2T-2.
METHODS: We conducted a short tandem repeat (STR) test to confirm the identity of the CBC2T-2 cell line. Furthermore, we assessed the migratory and invasive properties of the cells and performed clonogenicity assay to evaluate the ability of individual cells to form colonies. The tumorigenic potential of CBC2T-2 cells was tested in vivo using non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. The cells were injected subcutaneously and tumor formation was observed. In addition, immunohistochemical analysis was carried out to examine the expression of epithelial marker CK19 and mesenchymal marker vimentin in both CBC2T-2 cells and xenografts. The CBC2T-2 cell line was used to screen the potential therapeutic effects of various clinical agents in patients with cholangiocarcinoma sarcoma. Lastly, whole-exome sequencing was performed to identify genetic alterations and screen for somatic mutations in the CBC2T-2 cell line.
RESULTS: The STR test showed that there was no cross-contamination and the results were identical to those of the original tissue. The cells showed round or oval-shaped epithelioid cells and mesenchymal cells with spindle-shaped or elongated morphology. The cells exhibited a high proliferation ratio with a doubling time of 47.11 h. This cell line has migratory, invasive, and clonogenic abilities. The chromosomes in the CBC2T-2 cells were polyploidy, with numbers ranging from 69 to 79. The subcutaneous tumorigenic assay confirmed the in vivo tumorigenic ability of CBC2T-2 cells in NOD/SCID mice. CBC2T-2 cells and xenografts were positive for both the epithelial marker, CK19, and the mesenchymal marker, vimentin. These results suggest that CBC2T-2 cells may have both epithelial and mesenchymal characteristics. The cells were also used to screen clinical agents in patients with cholangiocarcinoma sarcoma, and a combination of paclitaxel and gemcitabine was found to be the most effective treatment option.
CONCLUSIONS: We established the first human cholangiocarcinoma sarcoma cell line, CBC2T-2, with stable biogenetic traits. This cell line, as a research model, has a high clinical value and would facilitate the understanding of the pathogenesis of cholangiocarcinoma sarcoma.
OBJECTIVE: To establish and characterize a new extrahepatic cholangiocarcinoma sarcoma cell line, namely CBC2T-2.
METHODS: We conducted a short tandem repeat (STR) test to confirm the identity of the CBC2T-2 cell line. Furthermore, we assessed the migratory and invasive properties of the cells and performed clonogenicity assay to evaluate the ability of individual cells to form colonies. The tumorigenic potential of CBC2T-2 cells was tested in vivo using non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. The cells were injected subcutaneously and tumor formation was observed. In addition, immunohistochemical analysis was carried out to examine the expression of epithelial marker CK19 and mesenchymal marker vimentin in both CBC2T-2 cells and xenografts. The CBC2T-2 cell line was used to screen the potential therapeutic effects of various clinical agents in patients with cholangiocarcinoma sarcoma. Lastly, whole-exome sequencing was performed to identify genetic alterations and screen for somatic mutations in the CBC2T-2 cell line.
RESULTS: The STR test showed that there was no cross-contamination and the results were identical to those of the original tissue. The cells showed round or oval-shaped epithelioid cells and mesenchymal cells with spindle-shaped or elongated morphology. The cells exhibited a high proliferation ratio with a doubling time of 47.11 h. This cell line has migratory, invasive, and clonogenic abilities. The chromosomes in the CBC2T-2 cells were polyploidy, with numbers ranging from 69 to 79. The subcutaneous tumorigenic assay confirmed the in vivo tumorigenic ability of CBC2T-2 cells in NOD/SCID mice. CBC2T-2 cells and xenografts were positive for both the epithelial marker, CK19, and the mesenchymal marker, vimentin. These results suggest that CBC2T-2 cells may have both epithelial and mesenchymal characteristics. The cells were also used to screen clinical agents in patients with cholangiocarcinoma sarcoma, and a combination of paclitaxel and gemcitabine was found to be the most effective treatment option.
CONCLUSIONS: We established the first human cholangiocarcinoma sarcoma cell line, CBC2T-2, with stable biogenetic traits. This cell line, as a research model, has a high clinical value and would facilitate the understanding of the pathogenesis of cholangiocarcinoma sarcoma.
摘要：
背景：肝外胆管癌肉瘤在临床上极为罕见。这些细胞由上皮细胞和间充质细胞组成。维持肿瘤特征的患者来源的细胞系是研究与癌肉瘤相关的分子机制的有价值的工具。然而,细胞库中没有胆管癌肉瘤细胞系。
目的：建立新的肝外胆管癌细胞，即CBC2T-2。
方法：我们进行了短串联重复（STR）测试，以确认CBC2T-2细胞系的身份。此外，我们评估了细胞的迁移和侵袭特性，并进行了克隆形成试验，以评估单个细胞形成集落的能力。使用非肥胖糖尿病/重度联合免疫缺陷(NOD/SCID)小鼠在体内测试CBC2T-2细胞的致瘤潜力。皮下注射细胞并观察肿瘤形成。此外,进行免疫组织化学分析以检查CBC2T-2细胞和异种移植物中上皮标志物CK19和间充质标志物波形蛋白的表达。CBC2T-2细胞系用于筛选各种临床药物对胆管癌肉瘤患者的潜在治疗作用。最后,在CBC2T-2细胞系中进行全外显子组测序以鉴定遗传改变并筛选体细胞突变.
结果：STR测试表明没有交叉污染，结果与原始组织相同。细胞呈圆形或椭圆形的上皮样细胞和间充质细胞，形态呈梭形或细长形。细胞表现出高增殖率，倍增时间为47.11h。该细胞系具有迁移，侵入性,和克隆能力。CBC2T-2细胞的染色体呈多倍体,数字从69到79不等。皮下致瘤测定证实了CBC2T-2细胞在NOD/SCID小鼠中的体内致瘤能力。CBC2T-2细胞和异种移植物对上皮标志物均呈阳性，CK19和间充质标志物，波形蛋白.这些结果表明CBC2T-2细胞可能同时具有上皮和间质特征。这些细胞还用于筛选胆管癌肉瘤患者的临床药物，紫杉醇和吉西他滨的联合治疗被认为是最有效的治疗选择。
结论：我们建立了第一个人胆管癌肉瘤细胞系，CBC2T-2,具有稳定的生物遗传性状。这个细胞系,作为一种研究模式，具有较高的临床应用价值,有助于了解胆管癌肉瘤的发病机制。
目的：建立新的肝外胆管癌细胞，即CBC2T-2。
方法：我们进行了短串联重复（STR）测试，以确认CBC2T-2细胞系的身份。此外，我们评估了细胞的迁移和侵袭特性，并进行了克隆形成试验，以评估单个细胞形成集落的能力。使用非肥胖糖尿病/重度联合免疫缺陷(NOD/SCID)小鼠在体内测试CBC2T-2细胞的致瘤潜力。皮下注射细胞并观察肿瘤形成。此外,进行免疫组织化学分析以检查CBC2T-2细胞和异种移植物中上皮标志物CK19和间充质标志物波形蛋白的表达。CBC2T-2细胞系用于筛选各种临床药物对胆管癌肉瘤患者的潜在治疗作用。最后,在CBC2T-2细胞系中进行全外显子组测序以鉴定遗传改变并筛选体细胞突变.
结果：STR测试表明没有交叉污染，结果与原始组织相同。细胞呈圆形或椭圆形的上皮样细胞和间充质细胞，形态呈梭形或细长形。细胞表现出高增殖率，倍增时间为47.11h。该细胞系具有迁移，侵入性,和克隆能力。CBC2T-2细胞的染色体呈多倍体,数字从69到79不等。皮下致瘤测定证实了CBC2T-2细胞在NOD/SCID小鼠中的体内致瘤能力。CBC2T-2细胞和异种移植物对上皮标志物均呈阳性，CK19和间充质标志物，波形蛋白.这些结果表明CBC2T-2细胞可能同时具有上皮和间质特征。这些细胞还用于筛选胆管癌肉瘤患者的临床药物，紫杉醇和吉西他滨的联合治疗被认为是最有效的治疗选择。
结论：我们建立了第一个人胆管癌肉瘤细胞系，CBC2T-2,具有稳定的生物遗传性状。这个细胞系,作为一种研究模式，具有较高的临床应用价值,有助于了解胆管癌肉瘤的发病机制。
