蛙皮素受体亚型-3在宫颈癌细胞能量代谢中的作用及机制探讨

Role and mechanism of fibrotin receptor subtype-3 (BRS-3) in energy metabolism of cervical cancer

  • 摘要:
    背景 宫颈癌是女性常见的恶性肿瘤,发病率在我国女性恶性肿瘤中居第二。
    目的 探讨蛙皮素受体亚型-3(Bombesin receptor subtype-3,BRS-3)在宫颈癌细胞能量代谢中的作用及机制。
    方法  将2021年6月— 2023年12月期间在新疆维吾尔自治区中医医院妇科门诊或住院的19例18 ~ 60岁之间的病理组织学确诊为宫颈癌患者作为宫颈癌组,选取同期门诊或住院治疗的18 ~ 60岁之间宫颈组织学确诊为良性病变的患者作为良性组。获得两组患者的宫颈组织标本。实时荧光定量PCR技术(quantitative real-time polymerase chain reaction,qRT-PCR)和免疫组化法检测人良性宫颈组织及人宫颈癌组织中蛋白表达水平和BRS-3、GSK3β、PRS6KB各基因表达水平。培养人宫颈癌细胞系Hela细胞,过表达或敲除BRS-3基因,Hela细胞随机分为空白对照组、空载对照组、BRS-3过表达组、阴性对照组和BRS-3沉默组,CCK-8检测Hela细胞增殖情况,检测Hela细胞乳酸、ATP、谷氨酸含量,采用qRT-PCR和Western blot方法检测BRS-3、GSK3β、RPS6KB1、PI3K和AKT mRNA和蛋白的表达水平。
    结果  与人良性病变宫颈组织相比,宫颈癌组织BRS-3 mRNA和蛋白表达水平均增高(P<0.05),GSK3β、PRS6KBmRNA表达水平增高(P<0.05)。培养Hela细胞,CCK-8检测结果显示,BRS-3过表达组Hela细胞存活率高于空白对照组、空载对照组(P<0.05);BRS-3沉默组Hela细胞存活率低于空白对照组、空载对照组、阴性对照组及BRS-3过表达组(P<0.05)。BRS-3过表达组Hela细胞中乳酸、ATP和谷氨酸含量高于空白对照组、空载对照组;BRS-3沉默组Hela细胞中乳酸、ATP和谷氨酸含量低于空白对照组、空载对照组、阴性对照组及BRS-3过表达组(P<0.05)。qRT-PCR检测结果显示,BRS-3沉默组中GSK3β、RPS6KB1、PI3K和AKT mRNA表达水平均低于空白对照组、阴性对照组(P<0.05)。Western blot实验结果显示,BRS-3沉默组BRS-3、GSK3β、RPS6KB1、PI3K和AKT蛋白表达表达水平以及p-AKT蛋白的磷酸化水平均低于空白对照组、阴性对照组(P<0.05)。
    结论  BRS-3在宫颈癌组织中表达升高,BRS-3可能通过增加Hela细胞有氧糖酵解促进肿瘤细胞增殖。

     

    Abstract:
    Background  Cervical cancer is a common malignant tumor in women, and its incidence rate ranks second among female malignant tumors in China.
    Objective To explore the role and mechanism of bombesin receptor subtype-3 (BRS-3) in the energy metabolism of cervical cancer cells.
    Methods Nineteen patients aged 18-60 years with pathologically confirmed cervical cancer who were treated in the Gynecology Outpatient Department or hospitalized in the Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous Region from June 2021 to December 2023 were selected as the cervical cancer group. Patients aged 18-60 years with histologically confirmed benign lesions of the cervix who were treated in the Outpatient Department or hospitalized during the same period were selected as the benign group. Cervical tissue specimens of the two groups of patients were obtained. Real-time fluorescence quantitative PCR and immunohistochemistry were used to detect the protein expression levels and the gene expression levels of BRS-3, GSK3β, and PRS6Kβ in human benign cervical tissues and human cervical cancer tissues. Human cervical cancer cell line Hela cells were cultured. The BRS-3 gene was overexpressed or knocked out. Hela cells were randomly divided into blank control group, empty vector control group, BRS-3 overexpression group, negative control group, and BRS-3 silencing group. CCK was used to detect the proliferation of Hela cells. Kits were used to detect lactate, ATP, and glutamate. Then, qRT-PCR and Western blot methods were used to detect the expression levels of target genes (GSK3β, RPS6Kβ1, PI3K, and AKT) and proteins (BRS-3, GSK3β, RPS6Kβ1, p-PI3K, AKT, and p-AKT).
    Results Compared with human benign cervical tissues, the expression of BRS-3 protein in cervical cancer tissues increased (P<0.05). Compared with human benign cervical tissues, the expression levels of BRS-3, GSK3β, and PRS6Kβ genes in cervical cancer tissues also increased (P<0.05). Human cervical cancer cell line Hela cells were cultured. qRT-PCR verified the overexpression efficiency of the BRS-3 gene. The expression of the BRS-3 gene in the BRS-3 overexpression group was higher than that in the blank control group and the empty vector control group (P<0.05). The results of cell proliferation detection showed that the survival rate of Hela cells in the BRS-3 overexpression group was higher than that in the blank control group and the empty vector control group (P<0.05). The survival rate of Hela cells in the BRS-3 silencing group was lower than that in the blank control group, the empty vector control group, the negative control group, and the BRS-3 overexpression group (P<0.05). The results of detecting the contents of lactate, ATP, and glutamate in Hela cells showed that the contents of lactate, ATP, and glutamate in the BRS-3 overexpression group were higher than those in the blank control group and the empty vector control group. The contents of lactate, ATP, and glutamate in the BRS-3 silencing group were lower than those in the blank control group, the empty vector control group, the negative control group, and the BRS-3 overexpression group (P<0.05). The results of qRT-PCR detection of the expression of each gene showed that the expression levels of GSK3β, RPS6Kβ1, PI3K, and AKT genes in the BRS-3 silencing group were lower than those in the blank control group and the negative control group (P<0.05). The results of Western blot showed that the expression levels of BRS-3, GSK3β, RPS6Kβ1, PI3K, and AKT proteins and the phosphorylation level of p-AKT protein in the BRS-3 silencing group were lower than those in the blank control group and the negative control group (P<0.05).
    Conclusion The expression of BRS-3 is increased in cervical cancer tissues. BRS-3 may promote tumor cell proliferation by increasing aerobic glycolysis of Hela cells.

     

/

返回文章
返回