Expression of Smo in pancreatic cancer CD44+CD24+cells and construction of a lentiviral expression vector to silence Smo

Abstract

The present study focused on the roles of members of the Hedgehog (Hh) signaling pathway in the maintenance of malignant biological characteristics, such as tumorigenesis, similar to that of pancreatic tumor cells. Cluster of differentiation (CD)44+CD24+/CD44−CD24− cells were isolated from three different pancreatic cancer cell lines by flow cytometry. Among the three pancreatic cancer cell lines, the SW1990 cell line exhibited the highest percentage of CD44+CD24+ cells, which accounted for 39.9% of the total. The expression of members of the Hh signaling pathway in CD44+CD24+/CD44−CD24− cells was detected using reverse transcription-polymerase chain reaction and western blot analysis. The results demonstrated that members of the Hh signaling pathway were differentially expressed in CD44+CD24+ cells compared with CD44−CD24−, normal pancreatic duct cells and unsorted SW1990 cells. In addition, lentiviral expression vectors expressing Smoothened (Smo) small interfering RNA (siRNA) were constructed. Following transfection with the lentiviral expression vectors, Smo expression was markedly reduced in CD44+CD24+ cells. The present study represents a preliminary investigation into the biological characteristics of CD44+CD24+ pancreatic cancer cells.

Resveratrol induces p53 in colorectal cancer through SET7/9

“…Resveratrol has been reported to be one of the most promising anticancer agents isolated from natural products that induce apoptosis in certain cancer types in a p53-dependent manner (17–20,32). Similarly, the present results propose resveratrol as a positive regulator of apoptosis in colorectal cancer cells (Fig.…”

“…The supernatant of medium was collected to infect the cancer cells. The viral titer was determined by Virus drops degree detection; fluorescence/absolute quantitative method (17). The cancer cells were infected by the collected supernatant for 48 h and collected for Western blot analysis.…”

“…p53 is mainly regulated through various post-translational modifications (PTMs), including acetylation, methylation, neddylation, phosphorylation and ubiquitination (13,16). Previous studies have demonstrated the involvement of resveratrol in the cellular function of p53 (17–20). Furthermore, the pro-apoptotic activity of resveratrol in prostate cancer appeared to be mediated by the serine-15 phosphorylation of p53 by mitogen-activated protein kinases (21).…”

<p>Knockdown of cyclooxygenase-2 leads to growth inhibition and cell cycle arrest in hepatocellular carcinoma cells</p>

“…Recently, lentivirus-mediated RNAi, which is mediated by 21–25 nucleotide short interfering RNA (siRNA) duplexes homologous to the interested gene, has rapidly become an innovative and elective tool for studying the regulation of gene expression and function in vitro 15. Furthermore, based on its high selectivity and specialty, this powerful tool is gradually become a novel anti-cancer therapeutic method by silencing some key oncogenic proteins 16.…”