具有高转移潜能的人肝癌细胞系的建立及其生物学特性

Establishment of a human hepatocellular carcinoma(HCC) cell line with high metastatic potential (MHCC97) and its biological characteristics

目的利用裸鼠人肝癌高转移模型（ＬＣＩ－Ｄ２０）的皮下移植瘤组织在体外建立一株具有高转移潜能的人肝癌细胞系（ＭＨＣＣ９７），并对其一般生物学特性进行观察。方法将分离的瘤细胞制成细胞悬液，用１０％人ＡＢ型血清的高糖ＤＭＥＭ培养液建成该细胞系，采用流式细胞术和染色体Ｇ－显带方法，进行细胞遗传学分析；用ＡＢＣ免疫组化法，观察其肺转移灶中癌细胞甲胎蛋白（ＡＦＰ）表达情况。结果ＭＨＣＣ９７细胞为典型的上皮样细胞，符合一般上皮性恶性肿瘤细胞的病理学特征。该细胞经皮下和肝内接种均可使裸鼠致瘤，并发生肺部转移。肝内接种者，肺转移达１００％（１２／１２）。ＭＨＣＣ９７细胞为异倍体细胞，染色体均为超二倍体，ｉ（１）（ｑ）和ｄｅｒ（４）（ｐｔｅｒ→ｑ３５：：？）等为其标志染色体，未显示有完整Ｙ染色体存在。肺转移灶的癌细胞ＡＦＰ阳性。结论ＭＨＣＣ９７细胞具有与原移植瘤相似的生物学特性。

Objective To establish and characterize a human hepatocellular carcinoma cell line with high metastatic potential derived from a subcutaneous xenograft of metastatic human HCC in nude mice (LCI D20). Methods Single cell suspension collected by tearing tumor tissues with forceps was transferred to the tissue culture flasks for culture in vitro in medium DMEM supplemented with 10% human group AB serum. Cytogenetic studies were performed on this cell line using flow cytometry and chromosome G banding. AFP of theprimary xenografts and lung metastatic lesions was detected by using ABC immunohistochemistry. The rates of its tumorigenicity and metastasis in nude mice were evaluated. Results The MHCC97 cells showed typical epithelial appearance. Upon subcutaneous or intrahepatic inoculation in nude mice, the xenograft grew and metastasized to the lungs. The metastatic rate was 100%. The cancer cells of lung metastatic foci were AFP positive. Aberrant chromosomes i(1)(q) and der(4) (pter→q35::?) were its chromosome markers. Conclusion The MHCC97 cell line maintained the biologic characteristics as its original xenografts. The presence of the reported chromosomal aberrations may be related to carcinogenesis and progression of HCC.