A comparative study of tumor-associated macrophages isolation in colorectal carcinoma by Ficoll-Hy-paque density gradient centrifugation and Percoll density gradient centrifugation / 实用医学杂志

Objective Percoll density gradient centrifugation and Ficoll-Hypaque density gradient cen-trifugation, which are frequently-used methods for separation of tumor-associated macrophages (TAMs) from solid carcinoma were compared, in order to find an effective way to separate TAMs from colorectal carcinoma (CRC). Furthermore, we studied the best adherence time of separating macrophage among mononuclear cells. Methods specimens were collected from CRC patients , after digesting into single cells , TAMs were separated from the same specimen by 100% Ficoll, 35% percoll and 25% combined with 65% percoll respectively. After these pre-liminary separation, the collected cells were purified a second time by adherence separation. The purity of TAMs were detected by immunofluorescence. Results TAMs purity from Ficoll-Hypaque density gradient centrifugation was 80.18%, statistically higher than that from Percoll density gradient centrifugations (54.33% and 10.93% re-spectively). Conclusion Compared to Percoll density gradient centrifugation, Ficoll-Hypaque density gradient centrifugation is a more effective and simple way to isolate TAMs from colorectal carcinoma , suggesting it can be wildly used in clinical and basic medical research. 2-4 hours is the best adherence time for isolating macrophage.

Expression of long noncoding RNA MALAT1 gene in human nasopharyngeal carcinoma cell lines and its biological significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the expression of MALAT 1 in nasopharyngeal carcinoma (NPC) and its biological function.</p><p><b>METHOD</b>Real-time PCR was used to detect the expression of MALAT1 in 5-8F, C666-1, CNE-1, CNE-2, HONE-1, 6-10B and NP69 cell lines. CNE-1 cells engineered with MALAT1 RNA interference (RNAi) and RNA activation (RNAa) techniques were examined for changes in cell proliferation, invasion and metastasis using CCK8 assay, colony formation assay, Transwell in vitro invasion assay and wound-healing assay ability.</p><p><b>RESULTS</b>MALAT1 was highly expressed in 5-8F cells with a high metastatic potential, and lowly expressed in normal nasopharyngeal epithelium cells. Overexpression of MALAT1 by RNAa suppressed the expression of E-cadherin, promoted the expression of vimentin and enhanced the proliferation, invasion, and metastasis of CNE-1 cells.</p><p><b>CONCLUSION</b>MALAT1 can enhance the proliferation, invasion, and metastasis of CNE-1 cells.</p>