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ObjectiveTo investigate the expression of lysophosphatidic acid （LPA） in patients with liver cancer， as well as its influence on malignant biological behavior of liver cancer and related regulatory mechanism. MethodsFrom January 2016 to December 2022， 26 patients with liver cancer， 28 patients with liver cirrhosis， and 28 individuals undergoing physical examination were enrolled. ELISIA was used to measure the content of LPA in plasma and peritoneal effusion of the patients with liver cancer or liver cirrhosis accompanied by peritoneal effusion， and the content of LPA was measured in plasma of the normal population at the same time， so as to clarify the difference in the expression of LPA in different populations， such as the patients with liver cancer and those with liver cirrhosis. MTT cell proliferation assay and cell migration assay were used to observe the influence of LPA and its inhibitor pertussis toxin （PTX） on the proliferation， migration， and invasion of SMMC7721 cells. In order to investigate the effect of LPA on the expression of RhoA and its upstream and downstream molecules FAK and P53 after binding to its receptor， qPCR and Western blot were used to observe the effect of LPA on the mRNA and protein expression levels of P53， FAK， and RhoA in SMMC7721 cells. A one-way analysis of variance was used for comparison of the means of continuous data between multiple groups， and the SNK-q test was used for comparison between two groups. ResultsCompared with the patients with liver cirrhosis， the patients with liver cancer had a significantly higher concentration of LPA in plasma （4.99±0.55 μmol/L vs 2.63±0.43 μmol/L， P<0.05） and peritoneal effusion （5.19±0.63 μmol/L vs 2.91±0.46 μmol/L， P<0.05）， and the patients with liver cancer also had a significantly higher level of plasma LPA than the normal population （4.99±0.55 μmol/L vs 1.61±0.39 μmol/L， P<0.05）. The cell proliferation assay showed that LPA significantly promoted the proliferation of SMMC7721 cells， and cell proliferation rate increased with the increase in dose and time； in particular， the middle-and high-dose groups had a significantly higher proliferation rate than the control group （P<0.05）. PTX inhibited the proliferative capacity of SMMC7721 cells in a time-dependent manner， and there was a significant difference between the groups （P<0.05）. The proliferation rate of the 72-hour high-dose LPA group was 3.6 times that of the control group， while the proliferation rate of the PTX group was 0.6 times that of the control group； the proliferation rate of the 72-hour high-dose LPA+PTX group was 1.2 times that of the control group. In addition， LPA increased the migration ability of hepatoma cells， while PTX inhibited their migration， in a time-dependent manner， and there was a significant difference between the groups （P<0.05）. The migration rate of the 72-hour high-dose LPA group was 3.09 times that of the control group， while the migration rate of the PTX group was 0.4 times that of the control group； the migration rate of the 72-hour high-dose LPA+PTX group was 0.99 times that of the control group. qPCR and Western blot showed that there were significant reductions in the mRNA and protein expression levels of P53 in SMMC7721 cells after LPA treatment， while there were significant increases in the mRNA and protein expression levels of FAK and RhoA； there was a significant difference between the LPA group and the control group （P<0.05）. ConclusionThere is an abnormal increase in the expression of LPA in patients with liver cancer， and LPA can promote the proliferation of liver cancer cells and increase their migration ability. At the same time， LPA changes the expression levels of P53， FAK， and RhoA， which may be associated with the promotion of tumor development and progression by LPA.
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Objective:To study the effect of vascular endothelial growth factor 165b (VEGF165b) on the biological characteristics of human hepatocellular carcinoma HepG2 cells,and to explore its mechanisr.Methods;The HepG2 cells were divided into blank group (treated with transfection reagent),control group (transfected with pcDNA3.0 expression vector) and pcDNA-VEGF165b group (transfected with pcNDA-VEGF165b).MTT assay was used to detect the survival rate of HepG2 cells;RT-PCR and Western blotting method were used to detect the expression levels of VEGF165b and VEGF165 mRNA and protein in the HepG2cells;Transwell chamber assay was used to measure the migration ability of HepG2 cells.Results:Compared with blank group,there was no significant changes in the expression levels of VEGF165b and VEGF165 mRNA and protein in the HepG2 cells in control group (P＞0.05).Compared with blank group,the expression levels of VEGF165b mRNA and protein in the HepG2 cells in PcDNA-VEGF165b group were significantly increased (P＜0.05),and the expression levels of VEGF165 mRNA and protein were significantly decreased (P＜0.05).There was no significant difference in the survival rates between blank group and control group (P＞0.05).The survival rate of HepG2 cells in PcDNA-VEGF165b group was lower than those in blank group and control group,but the differences were not statistically significant (P＞0.05).The cell migration experiment results showed that there was no significant difference in the migration rate between blank group and control group (P＜0.05).Compared with blank group and control group,the migration rate of HepG2 cells in PcDNA-VEGF165b group was significantly reduced (P＜ 0.05).Conclusion:VEGF165b can inhibit the expressions of VEGF165 mRNA and protein,and VEGF165b has no effect on the proliferation of hepatocellular carcinoma cells;but it can reduce the migration of hepatocellular carcinoma cells.
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Objective: To study the effect of vascular endothelial growth factor 165b (VEGF165b) on the biological characteristics of human hepatocellular carcinoma HepG2 cells, and to explore its mechanism. Methods: The HepG2 cells were divided into blank group (treated with transfection reagent), control group (transfected with pcDNA3. 0 expression vector) and pcDNA-VEGF165b group (transfected with pcNDA-VEGF165b). MTT assay was used to detect the survival rate of HepG2 cells; RT-PCR and Western blotting method were used to detect the expression levels of VEGF165b and VEGF165 mRNA and protein in the HepG2 cells; Transwell chamber assay was used to measure the migration ability of HepG2 cells. Results: Compared with blank group, there was no significant changes in the expression levels of VEGF165b and VEGF165 mRNA and protein in the HepG2 cells in control group (P>0. 05). Compared with blank group, the expression levels of VEGF165b mRNA and protein in the HepG2 cells in PcDNA-VEGF165b group were significantly increased (P0. 05). The survival rate of HepG2 cells in PcDNA-VEGF165b group was lower than those in blank group and control group, but the differences were not statistically significant (P>0. 05). The cell migration experiment results showed that there was no significant difference in the migration rate between blank group and control group (P<0. 05). Compared with blank group and control group, the migration rate of HepG2 cells in PcDNA-VEGF165b group was significantly reduced (P<0.05). Conclusion: VEGF165b can inhibit the expressions of VEGF165 mRNA and protein, and VEGF165b has no effect on the proliferation of hepatocellular carcinoma cells; but it can reduce the migration of hepatocellular carcinoma cells.
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Objective:To observe the effect of Allicin in cardial fibroblasts (CFs) proliferation and Collagen secretion,and to explore its role on TLR4/NF-κB signal pathway.Methods:CFs of neonatal Wistar rats were isolated and cultured ,then was stimulated with AngⅡ.CFs proliferation was measured by thiazolyl blue ( MTT) assay.The expression of collagenⅠ,collagenⅢ was measured by ELISA.mRNA expression of TLR4 and NF-κB were detected by reverse transcription-polymerase chain reaction ,protein expression of TLR4 and NF-κB were detected with Western blot.Results: Allicin could reduced MTT value of cardial fibroblasts ( P<0.01 ) , and inhibited expression of collagenⅠ,collagenⅢ(P<0.01),which in a dose-dependent manner.Allicin could reduced mRNA expression of TLR4 and NF-κB and protein expression of TLR 4 and NF-κB in CF induced by Ang Ⅱ ( all P<0.01 ) .Conclusion: Allicin can inhibit Myocardial fibrosis ,which mechanism is possible by inhibiting TLR 4/NF-κB signal pathway.
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Objective To explore the relation of the expressions of MDM2 and VEGF in osteosarcoma with the pathological parameters and prognosis of the tumor.Methods The expressions of MDM2 and VEGF were detected with immunohistochemical(SP) method in specimens from 56 cases of osteosarcoma.The correlation between the expressions of MDM2,VEGF and pathological grade,metastasis and prognosis was analyzed statistically.while 8 cases of fibrous dysplasia of bone were used as negative control group.Results The positive rates of MDM2 and VEGF in osteosarcoma were 64.3%(36/56) and 67.9%(38/56),respectively .MDM2 and VEGF didn't express in negative control group.The expression of MDM2 and VEGF were not significantly correlated to the pathological grades of the osteosarcoma,but which were significantly correlated with tumor metastasis and prognosis(P