SAMC在体内环境下通过干预MAPK与NF-κB信号通路发挥对乳腺癌的治疗作用

SAMC Treats Breast Cancer by Interfering with MAPK and NF-κB Signaling Pathway in Vivo

目的观察S-烯丙基巯基半胱氨酸(SAMC)在体内环境下对乳腺癌细胞MCF-7的抑制作用,并探讨其可能的分子机制。方法将乳腺癌细胞MCF-7接种于裸鼠左侧腋下,制作乳腺癌荷瘤裸鼠模型。将造模成功的裸鼠随机分为对照组、低剂量SAMC组与高剂量SAMC组。对照组给予等量生理盐水,低剂量SAMC组给予150 mg/kg的SAMC,高剂量SAMC组给予300 mg/kg的SAMC,各组每48 h腹腔注射给药1次,共计给药10次。①每4天测量一次各组裸鼠皮下肿瘤直径;②Q-PCR检测凋亡相关基因Bcl-2、Bax的转录水平;③Western Blot检测凋亡相关蛋白cleaved Caspase-3、Caspase-3、Bcl-2、Bax的表达水平;检测MAPK信号通路相关蛋白JNK、p-JNK、p38、p-p38的表达水平以及NF-κB信号通路相关蛋白p-IκBα、NF-κB p65的表达水平。结果①在相同时间点,对照组、低剂量SAMC组与高剂量SAMC组的瘤体直径依次递减(P<0.05);②Q-PCR结果显示:抗凋亡相关基因Bcl-2的转录水平降低,促凋亡相关基因Bax的转录水平升高;③Western Blot结果显示:凋亡相关蛋白cleaved Caspase-3、Bax的表达水平升高,而抗凋亡蛋白Bcl-2的表达水平降低(P<0.05);MAPK信号通路相关蛋白p-JNK、p-p38的蛋白表达水平升高(P<0.05),而对蛋白本身表达水平无明显影响(P>0.05);NF-κB信号通路中p-IκBα、p65的表达水平降低(P>0.05)。结论SAMC在体内环境下可以抑制荷瘤裸鼠的肿瘤生长,其机制可能与SAMC激活MAPK信号通路同时抑制NF-κB信号通路,从而促进肿瘤细胞凋亡有关。

Objective To observe the inhibitory effect of S-allyl-mercaptocysteine(SAMC)on MCF-7 in breast cancer cells and explore its possible molecular mechanism.Methods Breast cancer cells MCF-7 were inoculated into the left axilla of nude mice to make a nude mouse model of breast cancer.The nude mice that were successfully modeled were randomly divided into the control group,low-dose SAMC group and high-dose SAMC group.The control group was given the same amount of normal saline.The low-dose SAMC group was given 150 mg/kg SAMC,and the high-dose SAMC group was given 300 mg/kg SAMC.Each group was intraperitoneally injected once every 48 h for 10 times.①The diameter of subcutaneous tumors of nude mice in each group was measured every 4 days;②the transcription levels of apoptosis-related genes Bcl-2 and Bax were detected by Q-PCR;③The apoptosis-related protein cleaved Caspase-3 was detected by Western Blot,Caspase-3,Bcl-2,Bax expression levels;detection of MAPK signaling pathway-related proteins JNK,p-JNK,p38,p-p38 expression levels and NF-κB signaling pathway-related proteins p-IκBα,NF-κB p65 The level of expression.Results①At the same time point,the diameters of the tumors in the control group,the low-dose SAMC group and the high-dose SAMC group decreased in turn(P<0.05);②Q-PCR results showed that the anti-apoptosis-related gene Bcl-2 The transcription level decreased,and the transcriptional level of the apoptosis-promoting gene Bax increased.③Western Blot results show that the expression levels of apoptosis-related proteins cleaved Caspase-3 and Bax increased,while the anti-apoptotic protein Bcl-2 level decreased(P<0.05);the protein expression of p-JNK and p-p38 in MAPK signaling pathway increased(P<0.05),but had no significant effect on the expression of JNK and p38(P>0.05);The expression levels of p-IκBαand p65 in the NF-κB signaling pathway decreased(P>0.05).Conclusion SAMC inhibits the tumor growth in vivo,and the mechanism may be related to the activation of MAPK signaling pathway by SAMC and the inhibition of NF-κB signaling pathway,which promotes tumor cell apoptosis.