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Objective: To study the effects of oxymatrine and vincristine on resistance in HCT-8/VCR cells and its mechanism.
Methods: HCT-8 / VCR cells were cultured in vitro and were divided into blank control group, oxymatrine group, vincristine group, oxymatrine and vincristine combined group, each group had 6 complexes. The drug resistance of HCT-8/VCR cells was investigated by CCK-8 when treated with vincristine alone or in combination with oxymatrine. The autophagy was determined by monodansylcadaverine (MDC) staining. The level of IL-6 was detected by ELISA. The expressions of autophagy-related gene P62, LC3-Ⅱ / LC3-Ⅰ, Beclin-1 protein and TLR4 were detected by Western blot assay.
Results: Oxymatrine combined with vincristine could reduce the drug resistance of HCT-8 / VCR cells by the reversal multiple of 3.23. Compared with the blank control group, the content of autophagosome and the content of IL-6 in the oxymatrine group and the combination group were also decreased significantly (＜0.01). The content of autophagosome in the vincristine group was increased and the content of IL-6 was also significantly increased (＜0.01). Compared with the oxymatrine group, the combination group had higher autophagosome content, while IL-6 content was decreased (＜0.01); Western blot experiments showed that compared with the blank control group, the expression of P62 in the oxymatrine group was decreased (＜0.05), while the expressions of LC3-Ⅱ / LC3-Ⅰ, Beclin-1 and TLR4 were all increased (＜0.05). The expression of P62 in the vincristine group and the combined group was increased (＜0.05), and the expressions of LC3-Ⅱ / LC3-Ⅰ, Beclin-1, and TLR4 were all decreased (＜0.05). Compared with the vincristine group, the expression of P62 was increased in the combination group (＜0.05), and the expressions of LC3-Ⅱ / LC3-Ⅰ, Beclin-1, and TLR4 were all decreased (＜0.05).
Conclusion: Oxymatrine combined with vincristine can reduce the drug resistance of HCT-8/VCR cells, which may be related to the regulation of autophagy activity and TLR4 signal activation.
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| http://dx.doi.org/10.12047/j.cjap.5925.2020.075 | DOI Listing |
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