AAC-11 overexpression induces invasion and protects cervical cancer cells from apoptosis
AAC-11 overexpression induces invasion and protects cervical cancer cells from apoptosis
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DOI：
10.1038/labinvest.3780063
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发表时间：
2000-04-01
影响因子：
5
通讯作者：
Namkoong, SE
中科院分区：
文献类型：
--
作者：
Kim, JW;Cho, HS;Namkoong, SE
为了确定参与宫颈癌发生的基因，我们应用mRNA差异显示（DD）方法分析正常宫颈组织、宫颈癌、转移淋巴结和宫颈癌细胞系。我们克隆了一个491 bp的cDNA片段，CC 231，它存在于转移组织和宫颈癌细胞系中，但在正常宫颈和宫颈癌组织中不存在。491 bp的cDNA片段与先前发表的序列AAC-11（抗凋亡克隆11）具有98%的同源性。AAC-11 mRNA在9例正常宫颈组织和9例原发性宫颈癌组织中的表达水平均较低。其在三种转移组织和五种宫颈癌细胞系（HeLa、CaSki、SiHa、HKC-3和HKC-6）中的表达较高。AAC-11转染的AKC-6细胞对基质胶的侵袭和对层粘连蛋白的粘附分别增加了约2倍和4倍。北方印迹分析显示，基质金属蛋白酶（MMP）-2和1型膜型MMP（MT 1-MMP）基因在AAC-11转染的癌细胞中高水平表达。而MMP-2和MT 1-MMP在单独转染的细胞和野生型细胞中均不表达。AAC-11转染的细胞表达MMP-2蛋白的水平升高，通过免疫印迹评估。相反，MMP的组织抑制剂（TIMP-2）的表达在单独的载体转染的细胞或野生型细胞，分别检测。在AAC-11转染的细胞中未检测到其表达。在转染AAC-11的宫颈癌细胞中，β-catenin的表达上调。提示AAC-11转染导致MMP-2和MT 1-MMP过度表达、TIMP-2表达缺失以及β-catenin上调可能与宫颈癌侵袭的发展有关。AAC-11基因转染增加了宫颈癌细胞的定植。AAC-11对培养的宫颈癌细胞的作用与抗凋亡过程有关。无血清培养基中约50%的AAC-11转染细胞在2周后死亡，而单独载体或野生型细胞为1周。这些结果表明，AAC-11可能作为一个候选的转移相关基因和肿瘤抑制基因在人类宫颈癌。
To identify the genes involved in cervical carcinogenesis, we applied the mRNA differential display (DD) method to analyze normal cervical tissue, cervical cancer, metastatic lymph node, and cervical cancer cell line. We cloned a 491-bp cDNA fragment, CC231, which was present in metastatic tissue and cervical cancer cell line, but absent in normal cervical and cervical cancer tissues. The 491 bp cDNA fragment has 98% homology to the previously published sequence, AAC-11 (antiapoptosis clone 11). The levels of AAC-11 mRNA expressions in nine normal cervical and nine primary cervical cancer tissues were low. Its expression was higher in three metastatic tissues and five cervical cancer cell lines (HeLa, CaSki, SiHa, CUMC-3, and CUMC-6). Invasion of matrigel and adhesion to laminin by AAC-11 transfected CUMC-6 cells were increased by approximately 2-fold and 4-fold, respectively. Northern blot analysis showed that matrix metalloproteinase (MMP)-2 and membrane type 1 MMP (MT1-MMP) genes were found to be expressed in high levels in AAC-11-transfected cancer cells. But MMP-2 and MT1-MMP were not expressed in cells transfected with vector alone or wild-type cells. AAC-11-transfected cells expressed an elevated level of MMP-2 protein as assessed by immunoblotting. On the contrary, tissue inhibitor of MMP (TIMP-2) expression was detectable in cells transfected with vector alone or wild-type cells, respectively. Its expression was undetectable in AAC-11 transfected cells. In cervical cancer cells transfected with AAC-11, the expression of beta-catenin was up-regulated. These suggest that overexpressions of MMP-2 and MT1-MMP, loss of TIMP-2 expression, and up-regulation of beta-catenin by AAC-11 transfection may contribute to the development of cervical cancer invasion. AAC-11 gene transfection increased cervical cancer cell colonization. The effect of AAC-11 on cultured cervical cancer cells was associated with antiapoptotic process. Approximately 50% of the AAC-11 transfected cells in serum-free medium died after 2 weeks, compared to 1 week for vector alone or wild-type cells. These results suggest that AAC-11 may serve as a candidate metastasis-related and apoptosis-inhibiting gene in human cervical cancer.