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.