Mechanism of Polypeptide Extract from Scorpion Venom Combined Rapamycin in Enhancing Autophagy of H22 Hepatoma Cells: an Experimental Study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe enhanced effects of polypeptide extract from scorpion venom (PESV) combined Rapamycin on autophagy of H22 hepatoma cells in mice and to explore its possible mechanism.</p><p><b>METHODS</b>The H22 hepatocarcinoma cell suspension was subcutaneously inoculated into 40 Kunming mice. Then tumor-bearing mice were randomly divided into four groups, i.e., the control group,the high dose PESV group, the low dose PESV group, and the combination group (high dose PESV + Rapamycin), 10 in each group. Mice in high and dose PESV groups were administered with 20 mg/kg and 10 mg/kg PESV respectively by gastrogavage. Mice in the combination group were administered with 2 mg/kg rapamycin and 20 mg/kg PESV by gastrogavage. The intervention lasted for 14 successive days. The tumor volume was measured once every other day, the tumor growth curve was drawn, and then the tumor inhibitory rate calculated. Pathological changes of the tumor tissue were observed by HE staining. Protein expression levels of mammal target of rapamycin (mTOR), UNC-51-like kinase-1 (ULK1), microtubule-associated protein1 light chain3 (MAPILC3A), and Beclin1 were detected by immunohistochemical assay.</p><p><b>RESULTS</b>The growth of H22 hepatoma transplantation tumor was inhibited in high and low dose PESV groups and the combination group (P < 0.05). And there was statistical difference in tumor weight and tumor volume between the combination group and high and low dose PESV groups (P < 0.05). There was no statistical difference in tumor weight or tumor volume between the high dose PESV group and the low dose PESV group (P > 0.05). lmmunohistochemical assay showed that the protein expression of mTOR was higher, but protein expressions of ULK1, MAP1LC3A, Beclin1 were lower in the control group than in the rest 3 groups (P < 0.05, P < 0.01). Compared with the high dose PESV group, protein expressions of ULK1, MAP1LC3A, and Beclin1 were obviously lower (P < 0.05).</p><p><b>CONCLUSION</b>PESV combined Rapamycin might inhibit the development of H22 hepatoma transplantation tumor in mice possibly through inhibiting the activity of mTOR, enhancing expressions of ULK1, MAP1LC3A, and Beclin1.</p>

Study on the mechanism of polypeptide extract from scorpion venom on inhibition of angiogenesis of H 22 hepatoma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the mechanism of polypeptide extract from scorpion venom (PESV) on inhibiting angiogenesis.</p><p><b>METHODS</b>The H22 hepatoma tumor model was established by subcutaneously implanting H22 hepatoma cells into mice. The tumor-bearing mice were randomly divided into 4 groups, i.e., the control group, the high dose PESV group, the low dose PESV group, and the 5-fluorouracil (5-Fu) group, 10 mice in each group. The intervention was lasted for 14 days. The growth curve of the tumor volume was drawn and the inhibition rate calculated. Pathological changes of the tumors were observed by HE staining. The microvessel density (MVD) was detected using SP method. The protein expression levels of phosphatidylinositol 3-kinase (P13K), phosphoprotein kinase B (P-Akt), hypoxia-inducible factor-1 alpha (HIF-1 )alpha, and vascular endothelial growth factor-A (VEGF-A) were detected by immunohistochemical assay and Western blot.</p><p><b>RESULTS</b>The tumor inhibitory rate was 64.8%, 43.7%, and 32.4% in the 5-Fu group, the high dose PESV group, and the low dose PESV group. Compared with the control group, the protein expression of PI3K, P-Akt, HIF-1alpha, and VEGF-A were obviously inhibited by PESV and 5-Fu (P <0. 05,P <0. 01). The MVD also decreased in the high and low dose PESV groups (P < 0.05).</p><p><b>CONCLUSIONS</b>PESV could inhibit the angiogenesis of H22 hepatoma. The mechanisms might be associated with suppressing the expression of PI3K, P-Akt, HIF-1 alpha, and VEGF-A.</p>

Research on the effect and mechanism of polypeptide extract from scorpion venom combined with 5-fluorouracil on vasculogenic mimicry of H22 hepatoma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the inhibition effects of polypeptide extract from scorpion venom (PESV) combined 5-fluorouracil (5-Fu) on vasculogenic mimicry (VM) of H2 hepatoma carcinoma cells in mice and its possible mechanisms.</p><p><b>METHODS</b>The H22 carcinoma cell suspension was subcutaneously inoculated into 60 Kunming mice. Then tumor-bearing mice were randomly divided into three groups, i.e., the control group, the 5-Fu group, and the combination group (PESV +5-Fu), 20 in each group. The tumor volume was measured once every other day after 14 successive days of intervention. Then the tumor volume growth curve was drawn, and the tumor inhibitory rate was calculated. The morphological changes of the tumor tissue were observed by HE staining. The VM density of each tumor tissue were detected by immunohistochemical assay and periodic acid-schiff stain (PAS). The protein expression levels of hypoxia inducible factor-la (HIF-la) and matrix metalloproteinase-2 (MMP-2) were detected using immunohistochemical assay. The gray value was semi-quantitatively analyzed using LeicaQwinV3 Image Analysis Software.</p><p><b>RESULTS</b>The growth of H22 hepatoma transplantation tumor was inhibited more obviously in the combination group and the 5-Fu group than in the control group (P <0.05). There was statistical difference in the tumor weight and the tumor volume between the combination group and the 5-Fu group (P <0.05). Immunohistochemical assay and PAS showed that the VM density was obviously lower in the combination group than in the control group and the 5-Fu group (P <0.01). Compared with the control group, the protein expressions of HIF-la and MMP-2 significantly decreased in the combination group (P <0.01).</p><p><b>CONCLUSIONS</b>PESV combined 5-Fu could inhibit the generation of VM of H22 hepatoma transplantation tumor in mice. Its mechanisms might be associated with inhibiting the expressions of HIF-lalpha and MMP-2 in the microenvironment of tumors.</p>

Inhibition of polypeptide extract from scorpion venom on repopulation in Lewis lung adenocarcinoma during chemotherapy / 中草药

Objective: To identify the repopulation in metastatic Lewis lung adenocarcinoma during 5-Fu chemotherapy and observe the inhibition of polypeptide extract from scorpion venom (PESV) on the repopulation. Methods: To make repopulation model, Lewis cells were iv injected into caudal vein of C57BL/6 mice, the mice were ip injected by 5-Fu once every 7 d since day 7 and intervened by ig administration of PESV. Groups were treated differently. Six mice were sacrificed every 7 d, with counting metastatic foci in lung and detecting the level of proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF), and microvessel dentity (MVD) expression in Lewis lung metastatic loci by using immunohistochemistry. Results: In model groups, from day 21 to day 28 the numbers of metastatic big loci increased 2.5 averagely, while from day 14 to day 21 increased 0.8 only; The weight of lung from day 21 to day 28 increased more than that from day 14 to day 21; The expression of PCNA was the lowest in day 21, the highest in day 28, and there was no significant difference between day 28 and day 14; The expression in both high- and low-dose PESV groups in day 28 was lower than that in model group. There was a significant effect in high-dose PESV group. In model group the expression of VEGF in day 28 was upregulated significantly (P<0.01 vs that in model group in day 21). Compared with model group, VEGF expression in high-dose PESV group in day 21 and day 28 was downregulated, especially in day 28 (P<0.01). In low-dose PESV group only in day 28 the difference was found (P<0.05 vs that in model group); The change of MVD was the same as PCNA. The expression was the lowest in day 21, the highest in day 28, and there was no significant difference between day 28 and day 14. While in high-dose PESV group, the number of MVD in day 14 was more than that in day 21, and in day 21 more than in day 28, there were significant differences. In low-dose PESV group, in day 14 more than in day 21, but no significant difference was found between day 21 and day 28. Conclusion: The phenomenon of repopulation acceleration is found in Lewis lung adenocarcinoma during 5-Fu treatment and PESV could inhibit the repopulation through anti-angiogenesis which may be one of the mechanisms of inhibiting tumor cell repopulation.