Effects of aloperine on proliferation,apoptosis and immune escape of colorectal cancer cells by regulating IL-6/JAK1/STAT3 signaling pathway
Objective:To investigate the effects of aloperine(ALO)on cell behavior of colorectal cancer(CRC)cells through IL-6/tyrosine kinase 1(JAK1)/signal transducer and activator of transcription 3(STAT3)pathway.Methods:SW480 cells were grouped into CK group(normal culture of SW480 cells),ALO low-dose group(ALO-L group,0.2 mmol/L),ALO medium-dose group(ALO-M group,0.4 mmol/L),ALO high-dose group(ALO-H group,0.8 mmol/L)and ALO-H+activator(IL-6 activator recombinant human IL-6 protein)group(0.8 mmol/L+100 ng/ml).Proliferation of SW480 cells was detected by CCK-8 and plate cloning experi-ments;apoptosis of SW480 cells was detected by flow cytometry;Western blot was used to detect expressions of proliferating cell nu-clear antigen(PCNA),Bcl-2-associated X protein(Bax),IL-6,p-JAK1,p-STAT3 proteins in cells.After the above five groups of cells were co-cultured with natural killer cells NK-92MI,respectively,they were named as CK co-culture group,ALO-L co-culture group,ALO-M co-culture group,ALO-H co-culture group,and ALO-H+activator co-culture group,respectively.Levels of TNF-α and IFN-γ in supernatant and immune killing rate of NK-92MI in the co-culture system were detected.Results:Compared with CK group,OD450 value,clone formation rate,protein expressions of PCNA,IL-6,p-JAK1,p-STAT3 in SW480 cells in ALO-L group,ALO-M group and ALO-H group were decreased,while apoptosis rate and protein expression of Bax were increased,in a dose-dependent manner(P<0.05);compared with ALO-H group,OD450 value,clone formation rate,protein expressions of PCNA,IL-6,p-JAK1,p-STAT3 in SW480 cells in ALO-H+activator were increased,while apoptosis rate and protein expression of Bax were decreased(P<0.05);compared with CK co-culture group,levels of TNF-α and IFN-γ in supernatant of cells,and the immune killing rate of NK-92MI cells in ALO-L co-culture group,ALO-M co-culture group and ALO-H co-culture group were increased,and in a dose-dependent manner(P<0.05);compared with ALO-H co-culture group,levels of TNF-α and IFN-γ in supernatant of cells,and immune killing rate of NK-92MI cells in ALO-H+activator co-culture group were decreased(P<0.05).Conclusion:ALO may inhibit the proliferation,immune escape and promote apoptosis of SW480 cells by inhibiting IL-6/JAK1/STAT3 signaling pathway.
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