Intrinsic gene changes determine the successful establishment of stable renal cancer cell lines from tumor tissue

Abstract

Human tumor cell lines, especially those with complete data and follow-up, are important tools in tumor biological studies. Clear cell renal cell cancer (ccRCC) is not sensitive to radiotherapy or chemotherapy, and treatment of patients with distant metastasis relies on targeted therapy. Here, we report the establishment of seven new ccRCC stable cell lines that were continuously cultured for more than 20 generations among 81 cases of renal cell cancer. Moreover, gene expression and methylation in the established cell lines, in those that had a finite in vitro life span of less than 10 generations, and in cells that originated from the same culture at different generations were profiled using microarrays. Genes including SLC34A2, SEPP1, SULT1C4 and others were differentially expressed in established cell lines and finite cell lines, and changes in their expression might be caused by methylation or demethylation. The expression level of SLC34A2 was related not only to the life span in vitro culture but also to tumor size. Additionally, six of the seven new ccRCC cell lines had VHL deletions or termination mutations. So in addition to the establishment of seven new ccRCC cell lines with complete clinical data, we conclude that genes such as SLC34A2 and VHL play key roles in the continuous in vitro growth and development of ccRCC.

RNA sequencing reveals PNN and KCNQ1OT1 as predictive biomarkers of clinical outcome in stage III colorectal cancer patients treated with adjuvant chemotherapy

“…Furthermore, CYSLTR2 , cysteinyl leukotriene receptor 2, has been related to uveal melanoma progression . The upregulation of SULT1C4 would seem fundamental in maintaining establishment primary renal cell lines ccRCC . GIF , gastric intrinsic factor gene encodes a glycoprotein secreted by parietal cells of the gastric mucosa and is required for adequate absorption of vitamin B12.…”

LAG3(LAG-3,CD223) DNA methylation correlates with LAG3 expression by tumor and immune cells, immune cell infiltration, and overall survival in clear cell renal cell carcinoma

“…gov/ geo/, GSE82221, GSE89649). [17][18][19][20] Methylation analysis of the UHB cohort was performed using bisulfite-specific quantitative real-time PCR employing methylation-unspecific primers and probe pairs specifically and competitively binding methylated and unmethylated template DNA, respectively. This quantitative methylation-specific PCR (qMSP) is described in detail by Lehmann and Kreipe.…”

Effect of FAM196B in human lung adenocarcinoma

“…FAM196B is a protein coding gene and locates at 5q35.1. At present, it was only found that FAM196B was up-regulated in renal carcinoma 13 .DNA methylation of FAM196B showed high differences in breast cancer 18 .Recent studies showed that FAM196B expression was up-regulated in gastric cancer (GC) tissues and associated with poor tumor histology and T stage of GC. FAM196B also could promote gastric cancer cell proliferation through activating AKT signaling pathway 19 .…”

“…Chen and colleagues reported that FAM196B expressed higher in the embryonic ovary than the testis during mouse gonad development 12 . More recently, it was also reported that FAM196B was up-regulated in clear cell renal cell cancer cell line than finite cell lines 13 . Emerging evidence suggests FAM196B may participate in cancer development.…”