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  • Title: Cooperative effect of Bifidobacteria lipoteichoic acid combined with 5-fluorouracil on hepatoma-22 cells growth and apoptosis.
    Author: Guo B, Xie N, Wang Y.
    Journal: Bull Cancer; 2015 Mar; 102(3):204-12. PubMed ID: 25747889.
    Abstract:
    AIMS: To investigate the cooperative effect of Bifidobacteria lipoteichoic acid (BLTA) combined with 5-fluorouracil on tumor cells growth and apoptosis in mice bearing H22. METHODS: Hepatoma-22 (H22) cells were cultured in RPMI1640. Establish tumor-bearing mice model of liver cancer by injecting intraperitoneally 1×10(6)/mL cells into the above-mentioned Balb/c mice. 5-FU alone, BLTA alone or BLTA in combination with 5-FU were used to treat tumor-bearing mice. The tumor size were observed and measured regularly. The growth-inhibiting rate (IR) of tumor was detected. Real-Time PCR and Western blot were used to detect Bcl-2, Bax and Caspase-3 expressions of mRNA and protein in tumor tissue of tumor-bearing mice. Detection of apoptotic cells in tumor tissue by HE staining analysis. Detection of the organ index was for evaluate the added-activity of immune organs in mouse. FCM was used to detect T subgroup ratio of spleen cells of tumor-bearing mice. Expression change of mRNA and proteins of Foxp3 and TIM-3 were detected by Real-Time PCR and Western blot in tumor-bearing mice tumor tissue. RESULTS: BLTA and 5-FU significantly inhibited the proliferation of tumor and induced obvious apoptosis, the combined effects were greater than those of the individual agents (P<0.01). The underlying molecular mechanism of apoptotic process could be up-regulation of Bax and down-regulation of Bcl-2 and Caspase-3. The HE staining indicated that combined treat could both induce tissue cells necrosis and increase immune cells infiltration. Organ index showed that BLTA can enhance the proliferation of immune organs. The ratio of CD4(+)CD25(+) Treg significantly decreased and CD4(+) T cell increased in BLTA and 5-FU group (P<0.01). Compared to NS group, mRNA and proteins expression of Foxp3 and TIM-3 down regulated in BLTA and 5-FU group (P<0.01). CONCLUSIONS: These results show that combined effects of Bifidobacteria lipoteichoic acid and 5-FU on H22 cells were superior to the individual. The combination did not only increase anti-tumor effect, but also could alleviate the side effects of chemotherapy, with inhibiting TIM-3/TIM-3L pathway, cutting down immunosuppressive activity of CD4(+)CD25(+) Treg and enhancing cell-mediated immunity.
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