JAK2-STAT3通路介导牙龈卟啉单胞菌促结肠癌Caco-2细胞增殖的研究  

Porphyromonas gingivalis enhances the proliferation of colorectal cancer Caco⁃2 cells via the JAK2⁃STAT3 pathway

作　　者：张若彤 刘晓晨[2] 叶玮[1] ZHANG Ruotong;LIU Xiaochen;YE Wei(Department of Preventive Dentistry,Shanghai Ninth People’s Hospital,Shanghai Jiao Tong University School of Medicine,College of Stomatology,Shanghai Jiao Tong University,National Center for Stomatology,National Clinical Research Center for Oral Diseases,Shanghai Key Laboratory of Stomatology,Shanghai Research Institute of Stomatology,Shanghai 200011,China;Department of Gastroenterology,The Third Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110003,China)

机构地区：[1]上海交通大学医学院附属第九人民医院口腔预防科,上海交通大学口腔医学院,国家口腔医学中心,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海200011 [2]辽宁中医药大学附属第三医院消化内科,辽宁沈阳110003

出　　处：《口腔疾病防治》2023年第9期625-633,共9页Journal of Prevention and Treatment for Stomatological Diseases

基　　金：上海市科委港澳台科技合作项目(22410760200);上海市口腔疾病临床医学研究中心(19MC1910600);上海市临床重点专科项目(shslczdzk01601);上海市重中之重研究中心项目(2022ZZ01017)。

摘　　要：目的研究牙周致病菌—牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g)对人结肠癌Caco-2细胞增殖和炎症因子表达的影响,及JAK2-STAT3通路是否参与P.g对Caco-2细胞增殖的调控,为进一步探讨P.g与结肠癌之间的关系提供实验依据。方法体外培养Caco-2细胞,选择不同感染复数(multiplicity of infection,MOI)的P.g(0、1、10、25)刺激12、24、48 h,CCK8检测P.g对Caco-2细胞增殖的影响。设置刺激时间分别为12、24、48 h,MOI=0为对照组,MOI=1、10、25为实验组。qRT-PCR、Western blot检测各组白细胞介素-6(interleukin-6,IL-6)、白细胞介素-10(interleukin-10,IL-10)、信号转导与转录激活因子3(signal transducers and activators of transcription 3,STAT3)、蛋白酪氨酸激酶2(janus kinase 2,JAK2)基因和蛋白/磷酸化蛋白表达情况。结果P.g感染Caco-2细胞后,与对照组(MOI=0)相比,MOI=1和MOI=10时,P.g在12、24、48 h时对Caco-2细胞有持续刺激作用。与对照组相比,P.g感染Caco-2细胞中促炎因子IL-6及相关增殖通路因子STAT3、JAK2 mRNA表达及IL-6、p-STAT3、p-JAK2蛋白表达增加,且具有浓度和时间依赖性(P<0.05)。同时,P.g感染Caco-2细胞中抑炎因子IL-10的mRNA及蛋白表达降低(P<0.05)。添加JAK2抑制剂AZ960后,P.g感染Caco-2细胞的增殖减弱,STAT3、JAK2 mRNA表达及p-STAT3、p-JAK2蛋白表达降低(P<0.05)。结论P.g可促进结肠癌细胞系Caco-2的增殖,并且P.g作用于Caco-2细胞后可能通过JAK2-STAT3通路促进细胞增殖,同时促进促炎因子IL-6、抑制抑炎因子IL-10的表达,为细胞营造利于增殖的炎性环境,这可能是P.g影响Caco-2细胞增殖的机制之一。Objective To investigate the effect of pathogenic bacterium-Porphyromonas gingivalis(P.g)on the proliferation and inflammatory factor expression of human colorectal cancer Caco-2 cells,to determine whether the Janus kinase 2-signal transducers and activators of transcription 3(JAK2-STAT3)pathway is involved in the regulation of Caco-2 cell proliferation by P.g and to provide an experimental basis for further exploring the relationship between P.g and colorectal cancer.Methods Caco-2 cells were cultured in vitro,and P.g at different multiplicities of infection(MOIs)(0,1,10,25)was selected to stimulate for 12,24 and 48 h.The effect of P.g on the proliferation of Caco-2 cells was detected by CCK8.The stimulation time was set as 12,24 and 48 h.MOI=0 was the control group,and MOI=1,10 and 25 comprised the experimental group.qRT-PCR and Western blot were used to detect the changes in interleukin-6(IL-6),interleukin-10(IL-10),JAK2 and STAT3 gene and protein(phosphorylated protein)levels in each group.Results After P.g infection of Caco-2 cells,P.g had a sustained stimulatory effect on the cells for 12,24 and 48 h at MOI=1 and MOI=10 compared with the control group.Compared with that in the control group,the expression of pro-inflammatory factor IL-6 and related proliferative pathway protein JAK2 and STAT3 in Caco-2 cells with P.g infection increased in a concentration-and time-dependent manner(P<0.05).Additionally,the expression of IL-10,an anti-inflammatory factor,in Caco-2 cells infected with P.g decreased(P<0.05).After the addition of the JAK2 inhibitor AZ960,the proliferation of Caco-2 cells infected with P.g decreased,and the mRNA expression of STAT3 and JAK2 and the protein expression of p-STAT3 and p-JAK2 decreased(P<0.05).Conclusion P.g can promote the proliferation of the colorectal cancer cell line Caco-2,and the effect of P.g on Caco-2 cells may promote cell proliferation through the JAK2-STAT3 pathway while promoting the expression of the proinflammatory factor IL-6 and inhibiting the expression of the anti-

关 键 词：牙周炎 牙龈卟啉单胞菌 结肠癌细胞 白细胞介素⁃6 白细胞介素⁃10 蛋白酪氨酸激酶2 信号转导与转录激活因子3 

分 类 号：R78[医药卫生—口腔医学]