Effects of anti-heparanase antibody on the growth and invasion of HCCLM6 human hepatocellular carcinoma cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effects of the self-developed anti-heparanase polypeptide antibodies on growth and invasion of human hepatocellular carcinoma HCCLM6 cells.</p><p><b>METHODS</b>Using MTT, flow cytometry, plate clone formation, transwell invasion and heparan degrading enzyme assay, the growth and invasion changes of human hepatocellular carcinoma HCCLM6 cells by co-culture with each of three self-developed rabbit anti-heparanase polyclonal antibodies were detected.</p><p><b>RESULTS</b>Compared with normal rabbit IgG, in the presence of each anti-heparanase polypeptide antibody, the growth, cell cycle and clone formation remained unchanged, and under the P1 or P2 anti-heparanase polypeptide antibody (with final concentration 100 microg/ml), the cell invasiveness was inhibited by 52.5% and 36.6%, respectively, and the heparanase activity was inhibited by 42.9% and 39.1%, respectively.</p><p><b>CONCLUSION</b>The P1 and P2 anti-heparanase polypeptide antibodies can effectively inhibit the invasion ability and heparanase activity of liver cancer HCCLM6 cells. However, All the three antibodies have no effects on its growth, cell cycle and clone formation.</p>

Empirical study of oridonin-induced gastric cancer cells MKN45 apoptosis / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the growth inhibition and apoptosis of gastric cancer cell MKN45 induced by oridonin and its mechanism.</p><p><b>METHODS</b>The MTT method was used to investigate the inhibitory effect of oridonin on MKN45 cells. The AO/EB and Hoechst 33258 staining were used to observe the cell morphologic changes of apoptosis induced by oridonin. Prophase apoptotic ratio and cell cycle change were evaluated by GuavaEasycyte PCA-96 system. The expressions of Bcl-2, Bax and caspase 3 proteins were determined by Western blot.</p><p><b>RESULTS</b>Oridonin significantly inhibited the proliferation of MKN45 cells in dose- and time-dependent manner. Typical apoptotic features of the cells treated with oridonin were found by AO/EB and Hoechest33258 staining. When MKN45 cells were treated with different doses of oridonin for 12 h, the prophase apoptotic ratio was stepped up from 3.3% (untreated group) to 8.7%-17.9%; after 24 h, from 4.8% (untreated group) to 13.9%-29.3%. There was significant difference between treated and untreated groups (P <0.01). After treatment with oridonin for 24 h, MKN45 cells were arrested at G(2)/M phase. Western blot analysis showed up-regulated expression of Bax and caspase-3, and no significant change of Bcl-2, but Bcl-2/Bax ratio decreased significantly.</p><p><b>CONCLUSIONS</b>Oridonin significantly inhibits the proliferation of MKN45 cell. Apoptosis of MKN45 induced by oridonin may be associated with the up-regulated expression of Bax and the change of Bcl-2/Bax ratio, thus to activate the caspase pathway.</p>