Meng C, Li S L, Wang Y X, Fu J, Ge X Y
Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
Central Laboratory, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2024 Sep 9;59(9):942-949. doi: 10.3760/cma.j.cn112144-20240428-00171.
To establish a human buccal mucosa squamous cell carcinoma (BMSCC) cell line SCC117 in China, analyze and identify its basic biological characteristics. A 59-year-old Chinese male patient with BMSCC in the Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology in January 2011 was included in this study, his surgical specimens were primary cultured by improved tissue block culture method. The BMSCC cell line SCC117 was established after continuous passage and stable growth. The morphological characteristics of the cells were observed by light and electron microscope, and their basic biological characteristics were analyzed by growth curve, chromosome karyotype and xenotransplantation tumorigenicity in nude mice experiment. The expressions of cytokeratin (CK14), tumor-related proteins retinoblastoma tumor suppressor protein (RB), P53, E-cadherin, P21, phosphatase and tensin homolog deleted on chromosome ten (PTEN) were detected by immunohistochemical and human papilloma virus (HPV) were tested by PCR. SCC117 was identified by short tandem repeat (STR) analysis of genomic DNA. SCC117, a human BMSCC cell line, had been continuously subcultured for more than 150 generations. The cells grew in polygonal mosaic and lost contact inhibition, the typical desmosomes and tensional fibrils were observed by electron microscope, and CK14 was positive by immunohistochemistry. The doubling time was 40.16 h, the chromosome mode of the cell line was concentrated between 67 and 69, hypo-triploid. All 4 nude mice inoculated with SCC117 cells developed tumors, indicating that the SCC117 cell line had the ability of xenogeneic tumorigenesis. The histopathological type of the transplanted tumor in nude mice was consistent with that of the primary tumor tissue, both of which were squamous cell carcinoma. The immunohistochemical results showed that in both human primary tumor and the transplanted tumor tissue in nude mice, RB, P53, and E-cadherin were all positive, P21 was weakly positive, while PTEN was negative. SCC117 was tested negative for the presence of HPV. STR sequence analysis showed that SCC117 cell line originated from primary tumor tissue and was not cross-contaminated by other cell lines. The human BMSCC cell line SCC117 was successfully established in China, which could provide a new experimental model for the study of oral SCC without HPV infection, especially BMSCC.
在中国建立人颊黏膜鳞状细胞癌(BMSCC)细胞系SCC117,并分析鉴定其基本生物学特性。本研究纳入了一名2011年1月就诊于北京大学口腔医学院口腔颌面外科的59岁中国男性BMSCC患者,其手术标本采用改良组织块培养法进行原代培养。经连续传代和稳定生长后,建立了BMSCC细胞系SCC117。通过光镜和电镜观察细胞的形态特征,并通过生长曲线、染色体核型以及裸鼠异种移植致瘤性实验分析其基本生物学特性。采用免疫组织化学法检测细胞角蛋白(CK14)、肿瘤相关蛋白视网膜母细胞瘤抑癌蛋白(RB)、P53、E-钙黏蛋白、P21、第10号染色体缺失的磷酸酶和张力蛋白同源物(PTEN)的表达,采用PCR法检测人乳头瘤病毒(HPV)。通过对基因组DNA进行短串联重复序列(STR)分析鉴定SCC117。人BMSCC细胞系SCC117已连续传代150代以上。细胞呈多边形镶嵌生长,失去接触抑制,电镜下可见典型的桥粒和张力原纤维,免疫组织化学检测CK14呈阳性。倍增时间为40.16小时,该细胞系的染色体众数集中在67至69之间,为亚三倍体。接种SCC117细胞的4只裸鼠均长出肿瘤,表明SCC117细胞系具有异种致瘤能力。裸鼠移植瘤的组织病理学类型与原发肿瘤组织一致,均为鳞状细胞癌。免疫组织化学结果显示,在人原发性肿瘤和裸鼠移植瘤组织中,RB、P53和E-钙黏蛋白均呈阳性,P21弱阳性,而PTEN阴性。SCC117检测HPV呈阴性。STR序列分析表明,SCC117细胞系起源于原发肿瘤组织,未被其他细胞系交叉污染。在中国成功建立了人BMSCC细胞系SCC117,可为无HPV感染的口腔鳞状细胞癌尤其是BMSCC的研究提供新的实验模型。