Shi Xiuying, Jiang Biying, Liu Haifeng, Fan Chuifeng
Department of Pathology, First Affiliated Hospital and College of Basic Medical Sciences of China Medical University, Shenyang, China.
J Cell Biochem. 2019 Jun;120(6):10596-10604. doi: 10.1002/jcb.28346. Epub 2019 Jan 15.
ZCCHC9 is a type of CCHC type zinc-finger containing protein which was found to be expressed in some tissues including brain and testicles in mice. Expression and function of ZCCHC9 in human tissues including cancer was largely unknown. In this study, we investigated the expression and function of ZCCHC9 in human non-small cell lung cancer (NSCLC) and the related molecular mechanism. Immunochemistrical standing showed that ZCCHC9 was mainly located in the nucleus in bronchial epithelial cells and epithelial cells of submucosal glands (58.3% [14/24]). But in NSCLC cells ZCCHC9 was mainly located in the cytoplasm and the positive rate was 54.5% (60/110). Ectopic cytoplasmic expression of ZCCHC9 in cancer tissues was significantly associated with advanced TNM stages (III+IV), lymph node metastasis, and poor clinical outcome (P < 0.05). Overexpression of cytoplasmic ZCCHC9 using transfection of ZCCHC9 cDNA in A549 and NCI-H1299 cells significantly upregulated the proliferation and invasion of these cancer cells in vitro (P < 0.05). Western blot study showed that overexpression of cytoplasmic ZCCHC9 significantly upregulated expression of p-JNK, Cyclin D1, and MMP7 (P < 0.05). Next we used the inhibitor of JNK pathway to inhibit the activity of the JNK pathway and the results showed that co-addition of SP600125 significantly abolished the function of ZCCHC9 to promote the proliferation and invasion of cancer cells. These results indicate that cytoplasmic ZCCHC9 could promote the proliferation and invasion of NSCLC through the JNK pathway and may be a promising cancer maker.
ZCCHC9是一种含CCHC型锌指的蛋白质，在小鼠的一些组织（包括脑和睾丸）中被发现有表达。ZCCHC9在包括癌症在内的人体组织中的表达和功能在很大程度上尚不清楚。在本研究中，我们调查了ZCCHC9在人非小细胞肺癌（NSCLC）中的表达和功能以及相关分子机制。免疫组织化学染色显示，ZCCHC9主要位于支气管上皮细胞和黏膜下腺上皮细胞的细胞核中（58.3% [14/24]）。但在NSCLC细胞中，ZCCHC9主要位于细胞质中，阳性率为54.5%（60/110）。癌组织中ZCCHC9的异位细胞质表达与晚期TNM分期（III + IV期）、淋巴结转移及不良临床预后显著相关（P < 0.05）。通过在A549和NCI - H1299细胞中转染ZCCHC9 cDNA使细胞质ZCCHC9过表达，显著上调了这些癌细胞在体外的增殖和侵袭能力（P < 0.05）。蛋白质印迹研究表明，细胞质ZCCHC9过表达显著上调了p - JNK、细胞周期蛋白D1和基质金属蛋白酶7的表达（P < 0.05）。接下来我们使用JNK通路抑制剂抑制JNK通路的活性，结果显示，同时添加SP600125可显著消除ZCCHC9促进癌细胞增殖和侵袭的功能。这些结果表明，细胞质ZCCHC9可通过JNK通路促进NSCLC的增殖和侵袭，可能是一种有前景的癌症标志物。
