ABSTRACT
Objective To investigate the effect of EP4 gene silencing on the growth and migration of K1 cells. Methods K1 cells with stable knockdown of EP4 were constructed with lentiviral vector. QRT-PCR and western blot analysis were used to detect the expression of EP4 mRNA and protein in K1 cells. CCK8 assay and flow cytometry were employed to measure cell viability and apoptosis. Transwell assay was applied to detect cell migration. Results Compared with the negative control group,the mRNA and protein expression of EP4 were sig-nificantly decreased in K1 cells with stable knockdown of EP4. Furthermore,shRNA-mediated silencing of EP4 gene remarkably suppressed cell viability and induced apoptosis of K1 cells.The migration of K1 cells with knock-down of EP4 was decreased compared with the negative control group. Conclusions EP4 gene silencing can in-hibit growth and induce apoptosis of K1 cells.Downregulation of EP4 can significantly reduce migration of K1 cells.
ABSTRACT
@#The aim of the present study was to investigate the effects and possible mechanism of tetramethylpyrazine(TMP)on morphine-induced microglia activation and tolerance. The antinociception and morphine tolerance were assessed in mice using hot-water tail flick test. IBA-1(ionized calcium binding adapter molecule 1), the marker of microglia, was detected by immumofluorescence method. The expression of p-p38 MAPK and total p38 MAPK(mitogen-activated protein kinase, MAPK)was analyzed by Western blot; real-time polymerase chain reaction(RT-PCR)was used to detect the expression level of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β). Results showed that TMP(15, 30, 60 mg/kg, ip)inhibited morphine-induced up-regulation of IBA-1, p-p38, TNF-α and IL-1β in a dose-dependent manner, yet with no effect on the expression of total p38 MAPK. In conclusion, TMP significantly inhibited the activation of microglia evoked by morphine via p38 MAPK signaling pathway, thus attenuating morphine antinociception tolerance.
