Abstract 4966: Establishment and characterization of a paired human non-small cell lung cancer cell lines from primary cancer and metastasis lymph node

Abstract

Abstract Objective: Lung cancer has become one of the leading causes of cancer-related mortality in the world. Metastasis remains the overwhelming causes of death for lung cancer patients. 80-90% of lung cancer deaths are caused by metastasis. However, an ideal in vitro model to explore the regulation and the molecules that control the lymph node metastasis of lung cancer is required. Methods: A gene expression profiling for both of the cell lines were generated by microarray. Further investigations were carried out in the lung cancer cell lines and paired tissues by migration and invasion assays and immunohistochemistry. Results: Firstly, we established a paired human lung adenocarcinoma cell lines from primary tumor site (named DC cells) and metastatic paratracheal lymph nodes (named DL cells). The two cell lines have different biological characteristics which DC cells grow clump easily and DL cells grow more scattered. The doubling time of DC and DL cell lines were 32.6h and 45.2h, respectively. The detection of the human D1S80 and D17S30 VNTR locus verified the same individual for DC and DL cell lines. Secondary, Microarray analysis revealed 232 upregulated probe sets and 739 downregulated probe sets in DL cell line compared to DC cell line. Compared to DC cells, the transcriptional expression of 27 genes were dramatically increased (p < 0.05), including AIM2, IGFBP4, IL6 and IL6R in DL cells, while expression of 55 genes was clearly decreased (p < 0.05), including FGF13, FGF19, IL1A, IL1B, MCAM, MMP15, MMP7, and TGFB2 by real-time PCR analysis. Furthermore, we selected 32 genes and screened the expression of those genes in 10-paired human NSCLC primary lung cancer tissues and matched metastasis lymph nodes tissues by real-time PCR. The results showed: 8 out of 10 metastasis lymph node tissues got up-regulated of RGS7, VNN2, and 7 out of 10 metastasis lymph node tissues got up-regulated of PECAM1, VNN1, CLIC2 expression. On the other hand, 8 of 10 metastasis lymph node tissues got down-regulated expression of CEACAM6, and 7 out of 10 got down-regulated expression of PIAS3. At mean time, none of metastasis lymph node tissues got decreased expression of TNFRSF9, which only was seen in DL cells. The further study in other lung cancer cell lines indicated that CLIC2, PECAM1, RAB38, BCL9 and RGS7 genes influenc lung cancer cell migration ability in vitro. And these genes were also analyzed in more lung cancer tissues by immunohistochemistry staining. Conclusion: We established a paired human lung adenocarcinoma cell lines from primary tumor site and metastatic paratracheal lymph nodes. The CLIC2, PECAM1, RAB38, BCL9 and RGS7 genes involve in the lung cancer metastasis. Note: This abstract was not presented at the meeting. Citation Format: Jun Chen, Min Wang, Lingling Zu, Yongwen Li, Weiqiang Wang,Ying Li, Hongyu Liu. Establishment and characterization of a paired human non-small cell lung cancer cell lines from primary cancer and metastasis lymph node. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4966. doi:10.1158/1538-7445.AM2014-4966