Objective: To clarify the inhibitory mechanism of Haizao Yuhu Decoction on BRAFV600E mutation-driven papillary thyroid carcinoma cells. Methods: Prepare seaweed Yuhu Decoction medicated serum, select human normal thyr...Objective: To clarify the inhibitory mechanism of Haizao Yuhu Decoction on BRAFV600E mutation-driven papillary thyroid carcinoma cells. Methods: Prepare seaweed Yuhu Decoction medicated serum, select human normal thyroid cells Nthy-ori3-1 as the normal control group, and PLX4032 as the positive control drug. The experiment was divided into normal control group, model control group, PLX4032 group, Haizao Yuhu Decoction group (referred to as HYD group), Haizao Yuhu Decoction group + PLX4032 (referred to as HYD+PLX4032 group), and high iodine water group, at 8h and 24h respectively. At 72h, the cell proliferation of each group was detected by MTT method;at 24h, the expression of ERK and p-ERK protein in each group was detected by Western blot. Results: The longer the time, the more obvious the inhibitory effect of Haizao Yuhu Decoction-containing serum on the proliferation activity of BCPAP cells: Compared with the normal control group, the BCPAP cell proliferation activity of the model control group was significantly enhanced at 24h and 72h (P <0.05);At 24h, the PLX4032 group, HYD group, and HYD+PLX4032 group all showed a tendency to inhibit the proliferation of BCPAP cells, but there was no statistical difference;at 72h, compared with the model control group, the PLX4032 group The cell proliferation activity of the HYD group and HYD+PLX4032 group was significantly inhibited (P <0.05), and there was no difference between the high iodine water group and the model control group (P>0.05). In inhibiting the proliferation of BCPAP cells, the medicated serum of Shanghai Zaoyuhu Decoction has a synergistic effect with PLX4032 (F=10.87, P=0.005). Western blot results showed that there was no difference in the expression of ERK1/2 protein between the groups, but there were significant differences in the expression of p-ERK1/2 protein between the groups: Compared with the normal control group, the expression of p-ERK1/2 protein in the model group increased significantly (P < 0.05);Compared with the model control group, the expression of p-ERK1/2 protein in the PLX4032 group, the HYD group, and the HYD+PLX4032 group decreased significantly (P <0.05), and there was no difference between the high iodine water group and the model control group (P> 0.05). Conclusion: The medicated serum of Haizao Yuhu Decoction has an inhibitory effect on the proliferation of BCPAP cells, and its mechanism may be inhibiting the proliferation of BCPAP cells by inhibiting ERK1/2 protein phosphorylation, a protein post-translational modification process;the medicated serum of Haizao Yuhu Decoction may have Help enhance the effect of PLX4032 curative effect.展开更多
基金Fund Project:General Program of Natural Science Foundation of Liaoning Province(No.2015020390)General Program of National Natural Science Foundation of China(No.81874441)。
文摘Objective: To clarify the inhibitory mechanism of Haizao Yuhu Decoction on BRAFV600E mutation-driven papillary thyroid carcinoma cells. Methods: Prepare seaweed Yuhu Decoction medicated serum, select human normal thyroid cells Nthy-ori3-1 as the normal control group, and PLX4032 as the positive control drug. The experiment was divided into normal control group, model control group, PLX4032 group, Haizao Yuhu Decoction group (referred to as HYD group), Haizao Yuhu Decoction group + PLX4032 (referred to as HYD+PLX4032 group), and high iodine water group, at 8h and 24h respectively. At 72h, the cell proliferation of each group was detected by MTT method;at 24h, the expression of ERK and p-ERK protein in each group was detected by Western blot. Results: The longer the time, the more obvious the inhibitory effect of Haizao Yuhu Decoction-containing serum on the proliferation activity of BCPAP cells: Compared with the normal control group, the BCPAP cell proliferation activity of the model control group was significantly enhanced at 24h and 72h (P <0.05);At 24h, the PLX4032 group, HYD group, and HYD+PLX4032 group all showed a tendency to inhibit the proliferation of BCPAP cells, but there was no statistical difference;at 72h, compared with the model control group, the PLX4032 group The cell proliferation activity of the HYD group and HYD+PLX4032 group was significantly inhibited (P <0.05), and there was no difference between the high iodine water group and the model control group (P>0.05). In inhibiting the proliferation of BCPAP cells, the medicated serum of Shanghai Zaoyuhu Decoction has a synergistic effect with PLX4032 (F=10.87, P=0.005). Western blot results showed that there was no difference in the expression of ERK1/2 protein between the groups, but there were significant differences in the expression of p-ERK1/2 protein between the groups: Compared with the normal control group, the expression of p-ERK1/2 protein in the model group increased significantly (P < 0.05);Compared with the model control group, the expression of p-ERK1/2 protein in the PLX4032 group, the HYD group, and the HYD+PLX4032 group decreased significantly (P <0.05), and there was no difference between the high iodine water group and the model control group (P> 0.05). Conclusion: The medicated serum of Haizao Yuhu Decoction has an inhibitory effect on the proliferation of BCPAP cells, and its mechanism may be inhibiting the proliferation of BCPAP cells by inhibiting ERK1/2 protein phosphorylation, a protein post-translational modification process;the medicated serum of Haizao Yuhu Decoction may have Help enhance the effect of PLX4032 curative effect.