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目的目前结肠癌的治疗仍无突破性进展。本研究观察表达IL-24蛋白的重组双歧杆菌对大肠癌细胞NF-κB P65信号通路的抑制作用。方法采用分子克隆技术,先改建好在双歧杆菌高效表达的分泌型原核表达载体pBBADs,然后将人IL-24基因克隆到表达载体上,采用电穿孔法转化长双歧杆菌,获得携带IL-24基因的的重组双歧杆菌(BL-IL-24)。对BL-IL-24进行RT-PCR法行IL-24基因鉴定。结果 BL-IL-24的IL-24基因鉴定为621kb。鉴定最佳诱导表达时间为24h,BL-IL-24蛋白表达为21kb。BL-IL-24组和BL0组分别与PBS组,NF-κB p65、IL-6表达水平明显降低(P<0.05)。BL-IL-24与BL0组对比,NF-κB P65及IL-6表达水平降低(P<0.05)。结论 BL-IL-24的重组双歧杆菌结合了双歧杆菌及IL-24的双重治疗优势,通过抑制大肠癌细胞NF-κB P65及IL-6表达水平,从而,大肠癌有明显的治疗作用,为大肠癌的治疗提供新的治疗方法。
Objective The current treatment of colon cancer is still no breakthrough. This study was to observe the inhibitory effect of recombinant Bifidobacterium expressing IL-24 on NF-κB P65 signaling pathway in colorectal cancer cells. Methods The recombinant prokaryotic expression vector pBBADs was successfully constructed in Bifidobacterium. The human IL-24 gene was cloned into the expression vector and transformed into Bifidobacterium longum by electroporation to obtain the prokaryotic expression vector carrying IL- 24 gene of recombinant Bifidobacterium (BL-IL-24). IL-24 gene was identified by RT-PCR on BL-IL-24. Results The IL-24 gene of BL-IL-24 was identified as 621 kb. Identification of the best induction of expression for 24h, BL-IL-24 protein expression of 21kb. The levels of NF-κB p65 and IL-6 in BL-IL-24 group and BL0 group were significantly lower than those in PBS group (P <0.05). The expression of NF-κB P65 and IL-6 in BL-IL-24 group was lower than that in BL0 group (P <0.05). Conclusion The recombinant Bifidobacterium of BL-IL-24 binds the dual therapeutic advantages of Bifidobacterium and IL-24 and has a significant therapeutic effect on colorectal cancer by inhibiting the expression of NF-κB P65 and IL-6 in colorectal cancer cells , For the treatment of colorectal cancer provides a new treatment.