| Objective:This study to detect the prevalence of Toxoplasma gondii infection among patients with malignant tumors in Dali City,so as to further enrich the database of T.gondii infection and serve as a guide for the prevention and control of T.gondii infection in patients with malignant tumors.At the same time,in order to provide new thoughts for the therapeutic treatment of malignant tumors,the effects of the culture supernatant of T.gondii on the proliferation,apoptosis,migration,and invasion of malignant tumor cells were also examined.Methods:1.The serum samples of 721 patients with malignant tumors treated at the First Affiliated Hospital of Dali University between 2020 and 2022 were collected,and the serum of 100 healthy medical examiners was also collected as controls.The enzyme-linked immunosorbent assay was used to detect anti-Toxoplasma gondii IgG and IgM antibodies in serum,and to compare and analyze the differences in the positive rates of anti-Toxoplasma gondii specific antibodies in patients with different types and characteristics of malignant tumors.2.The cell culture plate was inoculated with bladder cancer 5637 cells at logarithmic phase,and the supernatant of RH type and ToxoDB#9 type Toxoplasma gondii after 24 h of culture was added to the experimental group,and an equal volume of RPMI1640 medium was added to the control group.The morphological changes of cells after Toxoplasma culture supernatant intervention were observed by microscopy,the effect of Toxoplasma culture supernatant on cell proliferation ability was detected by CCK-8 experiment,the effect on migration and invasion ability of cells was detected by using scratch healing experiment and Transwell experiment,and the effect on apoptosis rate of cells was detected by using TUNEL experiment.The effect on the expression of apoptosis-related proteins Bax and Bcl-2 in the cells was examined by Western blot experiment.Statistical analysis of the experimental results was performed using statistical software to draw valid conclusions.Result:1.The positive rate of serum anti-Toxoplasma IgG antibody was 21.22% in 721 patients with malignant tumors,which was higher than that of 8.00% in healthy controls(P<0.05),and the difference between the positive rate of serum anti-Toxoplasma IgM antibody and that of healthy controls was not statistically significant(P>0.05).Among different types of malignant tumors,the positive rates of serum anti-Toxoplasma IgG antibodies in patients with gastric cancer,bladder cancer,colorectal cancer,thyroid cancer,lung cancer,cervical and endometrial cancer and liver cancer were significantly higher than those in healthy controls(P < 0.05),and the positive rates of serum anti-Toxoplasma IgM antibodies in patients with cervical and endometrial cancer were significantly higher than those in healthy controls(P<0.05),but the positive rates of serum anti-Toxoplasma IgM antibodies in patients with different types of malignant tumors.However,there was no statistically significant difference in the positive rates of anti-Toxoplasma IgG and IgM antibodies among patients with different types of malignant tumors(P > 0.05).The differences in serum anti-Toxoplasma gondii IgG antibody positivity rates among patients with malignant tumors of different gender,ages,ethnicities,occupations and places of residence were not statistically significant(P>0.05).The differences in serum anti-Toxoplasma gondii IgM antibody positivity rates among patients with malignant tumors of different gender,ages were not statistically significant(P<0.05).2.(1)The cell morphology was observed microscopically.After 24 h of Toxoplasma culture supernatant intervention,the RH group and ToxoDB#9 group showed reduced cell volume,cell crinkling,reduced nuclear plasma ratio,pyknosis and rupture compared to the control group.(2)In the CCK-8 experiment,Toxoplasma gondii culture supernatant had an inhibitory effect on the proliferative capacity of 5637 cells,and the inhibition rate was higher in the ToxoDB#9 group than in the RH group(P<0.05).(3)In the scratch healing experiment,the cell migration rates of the control,RH and ToxoDB#9 groups at 24 h were(19.92±8.62)%,(11.80±3.18)%,and(4.02±1.73)%.The cell migration rates of the control,RH and ToxoDB#9 groups at 36 h were(64.60±5.60)%,(17.57±4.24)%,and(15.47±4.82)%.Compared with the control group,the cell migration rate was reduced in both RH and ToxoDB#9 groups(P<0.05),and the migration rate was lower in the ToxoDB#9 group than in the RH group(P<0.05).(4)In the Transwell migration experiment,the number of migrated cells in the control,RH and ToxoDB#9 groups were(899.33±28.19),(476.00±25.96)and(439.00±31.38).Compared with the control group,the number of migrated cells was lower in both the RH and ToxoDB#9 groups(P<0.05),and the number of migrated cells was lower in the ToxoDB#9 group than in the RH group(P>0.05).(5)In the Transwell invasion experiment,the number of invasive cells in the control,RH and ToxoDB#9 groups were(459.67±11.61),(343.33±20.74)and(217.33±32.42).Compared with the control group,the number of invasive cells was lower in both the RH and ToxoDB#9 groups(P<0.05),and the number of invasive cells was lower in the ToxoDB#9 group than in the RH group(P<0.05).(6)In the TUNEL experiment,the apoptosis rates of the control,RH and ToxoDB#9 groups were(5.71±2.37)%,(74.35±5.88)% and(79.29±6.54)%.Compared with the control group,the apoptosis rate increased in both RH and ToxoDB#9 groups(P<0.05),and the apoptosis rate in ToxoDB#9 group was higher than that in RH group(P>0.05).(7)In the Western blot experiment,the Bax protein expression level was significantly increased and the Bcl-2 protein expression level was significantly decreased in the cells of RH group and ToxoDB#9 group compared with the control group,and the Bcl-2/Bax ratio was decreased,and the difference was statistically significant as shown by statistical analysis(P<0.05).Conclusion:1.The infection rate of Toxoplasma gondii infection in patients with malignant tumors in Dali was significantly higher than that in healthy controls,but the difference in the infection rate of Toxoplasma gondii among patients with different types of malignant tumors was not statistically significant.The infection rate of Toxoplasma gondii in patients with malignant tumors was not specific for gender,age,occupation,place of residence and ethnicity.It is suggested that the combination of Toxoplasma gondii infection in malignant tumor patients is relatively common,and the prevention and treatment of Toxoplasma gondii infection should be strengthened during clinical treatment.2.Toxoplasma gondii culture supernatant significantly inhibited the proliferation,migration and invasion ability of bladder cancer 5637 cells,while Toxoplasma gondii culture supernatant promoted apoptosis of cells by upregulating Bax protein and downregulating Bcl-2 protein expression.The inhibitory effect of ToxoDB#9 strain culture supernatant on bladder cancer 5637 cells was stronger than that of RH strain culture supernatant under the same time intervention. |
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