Low Doses of Nonylphenol Promote Growth of Colon Cancer Cells through Activation of ERK1/2 via G Protein‒Coupled Receptor 30 / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Nonylphenol (NP) is an endocrine disruptor found in products such as cleaners, plastics, and detergents. It exerts actions similar to endogenous 17β-estradiol (E2) and is reported to influence various cancers. However, its role in colon cancer remains elusive. MATERIALS AND METHODS: Colon cancer cell lines COLO 205 and SW480 were employed in our study. The cells were treated with NP or E2 followed by measurement of apoptosis and proliferation using flow cytometry and MTT assays, respectively. G protein–coupled estrogen receptor 30 (GPR30) expression was visualized using immunofluorescence and Western blot. To investigate the underlying mechanism, the expression levels of GPR30, p-protein kinase A (PKA), c-myc, cyclin D1, and ERK1/2 were analyzed using Western blot. Meanwhile, the GPR30 antagonist G15 was utilized to validate the role of GPR30 in colon cancer progression. Finally, the effect of a GPR30 inhibitor on tumor growth was determined in vivo using tumor xenograft mouse models. RESULTS: NP facilitated the proliferation of colon cancer cells and induced apoptosis failure in vitro. Western blot revealed increased GPR30 expression levels in response to NP treatment. Cyclin D1, p-PKA, c-myc, and proliferating cell nuclear antigen, proteins that regulate the cell cycle, were all upregulated by NP, and NP-mediated ERK1/2 activation and subsequent cell proliferation were abrogated by the GPR30 inhibitor G15. Moreover, colon cancer mice that received G15 administration demonstrated impaired tumor growth in vivo. CONCLUSION: Low dose NP promotes the growth of colon tumors through GPR30-mediated activation of ERK1/2 signaling.

Genistein Inhibits Proliferation of BRCA1 Mutated Breast Cancer Cells: The GPR30-Akt Axis as a Potential Target

BACKGROUND: BRCA1 mutated breast cancer cells exhibit the elevated cell proliferation and the higher metastatic potential. G protein-coupled receptor 30 (GPR30) has been shown to regulate growth of hormonally responsive cancers, such as ovarian and breast cancers, and high expression of GPR30 is found in estrogen receptor (ER)-negative breast cancer cells. ER-negative breast cancer patients often have a mutation in the tumor suppressor gene, BRCA1. This study explored antiproliferative effects of genistein, a chemopreventive isoflavone present in legumes, and underlying molecular mechanisms in triple negative breast cancer cells with or without functionally active BRCA1.METHODS: Expression of BRCA1, GPR30 and Nrf2 was measured by Western blot analysis. Reactive oxygen species (ROS) accumulation was monitored by using the fluorescence-generating probe, 2’,7’-dichlorofluorescein diacetate. The effects of genistein on breast cancer cell viability and proliferation were assessed by the MTT, migration and clonogenic assays.RESULTS: The expression of GPR30 was dramatically elevated at both transcriptional and translational levels in BRCA1 mutated breast cancer cells compared to cells with wild-type BRCA1. Notably, there was diminished Akt phosporylation in GPR30 silenced cells. Treatment of BRCA1 silenced breast cancer cells with genistein resulted in the down-regulation of GPR30 expression and the inhibition of Akt phosphorylation as well as the reduced cell viability, migration and colony formation. Genistein caused cell cycle arrest at the G₂/M phase in BRCA1-mutant cells through down-regulation of cyclin B1 expression. Furthermore, BRCA1-mutant breast cancer cells exhibited higher levels of intracellular ROS than those in the wild-type cells. Genistein treatment lowered the ROS levels through up-regulation of Nrf2 expression.CONCLUSIONS: Lack of functional BRCA1 activates GPR30 signaling, thereby stimulating Akt phosphorylation and cell proliferation. Genistein induces G2/M phase arrest by down-regulating cyclin B1 expression, which is attributable to its suppression of GPR30 activation and Akt phosphorylation in BRCA1 impaired breast cancer cells.

Effects of estrogen receptor GPR30 agonist G1 on neuronal apoptosis and microglia polarization in traumatic brain injury rats / 中华创伤杂志(英文版)

<p><b>PURPOSE</b>To investigate the effects of estrogen G protein-coupled receptor 30 (GPR30) agonist G1 on hippocampal neuronal apoptosis and microglial polarization in rat traumatic brain injury (TBI).</p><p><b>METHODS</b>Male SD rats were randomly divided into sham group, TBI + vehicle group, TBI + G1 group. Experimental moderate TBI was induced using Feeney's weigh-drop method. G1 (100μg/kg) or vehicle was intravenously injected from femoral vein at 30 min post-injury. Rats were sacrificed at 24 h after injury for detection of neuronal apoptosis and microglia polarization. Neuronal apoptosis was assayed by immunofluorescent staining of active caspase-3. M1 type microglia markers (iNOS and IL-1β) and M2 type markers (Arg1 and IL-4) were examined by immunoblotting or ELISA. Total protein level of Akt and phosphorylated Akt were assayed by immunoblotting.</p><p><b>RESULTS</b>G1 significantly reduced active caspase-3 positive neurons in hippocampus. Meanwhile G1 increased the ratio of Arg1/iNOS. IL-1β production was decreased but IL-4 was increased after G1 treatment. G1 treatment also increased the active form of Akt.</p><p><b>CONCLUSIONS</b>GPR30 agonist G1 inhibited neuronal apoptosis and favored microglia polarization to M2 type.</p>

Differential expression of estrogen receptor GPR30 in main supporting ligaments of patients with pelvic organ prolapsed / 上海交通大学学报（医学版）

Objective · To investigate the expression levels of estrogen receptor GPR30, elastin, collagen Ⅰ (Col Ⅰ ), and Col Ⅲ in the uterus round ligament and the cardial ligament in patients with pelvic organ prolapse (POP) and main components in extracellular matrix (ECM). Methods · 22 patients with POP (the POP group) and 10 patients with cervical intraepithelial neoplasia (CIN) Ⅲ (the control group) were enrolled. Hysterectomy specimens were collected and expressions of GPR30, elastin, Col Ⅰ and Col Ⅲ in the uterine round ligament and the cardial ligament tissues were detected with Western blotting and immunohistochemistry. Results · Western blotting and immunohistochemistry results showed that the expressions of GPR30, elastin, Col Ⅰ and Col Ⅲ in the uterine round ligament and the cardial ligament tissues were significantly lower in the POP group than in the control group. The differences were statistically significant (P<0.05). Conclusion · The occurrence of POP may be associated with estrogen receptor deficiency and decreased contents of collagen and elastin in relevant tissues. Whether GPR30 is involved in the synthesis, secretion and degradation of ECM major components in pelvic support tissues needs further investigation.

Differential expression of estrogen receptor GPR30 in main supporting ligaments of patients with pelvic organ prolapsed / 上海交通大学学报（医学版）

Objective · To investigate the expression levels of estrogen receptor GPR30, elastin, collagen Ⅰ (Col Ⅰ ), and Col Ⅲ in the uterus round ligament and the cardial ligament in patients with pelvic organ prolapse (POP) and main components in extracellular matrix (ECM). Methods · 22 patients with POP (the POP group) and 10 patients with cervical intraepithelial neoplasia (CIN) Ⅲ (the control group) were enrolled. Hysterectomy specimens were collected and expressions of GPR30, elastin, Col Ⅰ and Col Ⅲ in the uterine round ligament and the cardial ligament tissues were detected with Western blotting and immunohistochemistry. Results · Western blotting and immunohistochemistry results showed that the expressions of GPR30, elastin, Col Ⅰ and Col Ⅲ in the uterine round ligament and the cardial ligament tissues were significantly lower in the POP group than in the control group. The differences were statistically significant (P<0.05). Conclusion · The occurrence of POP may be associated with estrogen receptor deficiency and decreased contents of collagen and elastin in relevant tissues. Whether GPR30 is involved in the synthesis, secretion and degradation of ECM major components in pelvic support tissues needs further investigation.

Expression of GPR30,HRG1 and HER2 in breast cancer and their relationship with lymphatic metastasis / 重庆医学

Objective To explore the expression of GPR30,HRG1 and HER2 including the activation status of HER2 (phosphorylated HER2)in invasive ductal breast cancers and their relationship with lymphatic metastasis.Methods The expres-sion of GPR30,HRG1,HER2 and pHER2 in 72 cases of specimens of invasive ductal breast cancers were examined by immunohis-tochemistry method.Results A moderate correlation between GPR30 and HRG1 was disclosed (r=0.597,P =0.000).There was strong correlation between pHER2 and GPR30 or HRG1(r=0.742,P =0.000;r=0.615,P =0.000).The expression of GPR30 and pHER2 in the lymphatic metastasis group was remarkably higher than in the group without lymphatic metastasis(P <0.05). Conclusion The interaction between GPR30 and HRG1 HER2 signal transduction pathways might be involved in the lymphatic metastasis in breast cancer.Blocking both of GPR30 and HRG1 signaling pathway could be a promising new strategy for breast cancer treatments.

Expression and significance of Caveolin-1, GPR30 and Vimentin in papillary thyroid carcinoma / 中国免疫学杂志

Objective:To investigate the expression of Caveolin-1,GPR30 and Vimentin at tumor tissues of human papillary thyroid carcinoma( PTC) patients and analyze their associations for the possible clinical implication.Methods: Immunohistochemistry was used to analyze the expression of Caveolin-1, GPR30 and Vimentin in 76 PTC and 44 normal samples.The correlations of Caveolin-1, GPR30 and Vimentin expression with one another, and with several clinicopathological indicators were statistically analyzed.Results:In PTCs,the positive expression rates of Caveolin-1,GPR30 and Vimentin were 9.21%(7/76),80.26%(61/76) and 76.32%(58/76),respectively.Caveolin-1,GPR30 and Vimentin expression had significant correlations with TNM stages(P=0.005,P<0.001,and P<0.001,respectively) and with cervical lymph node metastasis(P≤0.001 for all).Meanwhile,Caveolin-1 ex-pression had a negative correlation with GPR30(rs=-0.528,P<0.001) and Vimentin(rs=-0.572,P<0.001).GPR30 and Vimentin expression were positively correlated ( rs=0.812, P<0.001 ).Caveolin-1 under-expression was accompanied by GPR30 or Vimentin over-expression had stronger correlation with LNM ( P=0.020 for Caveolin-1/GPR30 and P=0.001 Caveolin-1/Vimentin ) than did each alone;concomitant high expression of GPR30 and Vimentin had stronger correlation with LNM( P=0.005 for GPR30/Vimentin) than did each alone.And lower expression of Caveolin-1 accompanied by higher expression of GPR30 and Vimentin was significantly as-sociated with LNM as compared with cases not showing such expressing(P<0.001).Conclusion:These results demonstrated that the evaluation of Caveolin-1 ,GPR30 and Vimentin expression may be play an important role in the development and metastasis of PTC,and their biological function may relate with each other.

The Effects of Nonylphenol on Calcium Signaling and Cell Proliferation in H9c2 Cell and Its Mechanism / 中国医科大学学报

Objective To observe the effects of nonylphenol(NP)on the intracellular calcium concentration changes and cell proliferation,and the involvement of GPR30 receptor in H9c2 cell. Methods The intracellular calcium concentration changes were recorded by using intracellular calcium determination method and cell proliferation was observed by MTT method in H9c2 cell. Results NP(1×10-10 mol/L)increased the intra?cellular calcium concentration changing amplitude and promoted the proliferation of H9c2 cells,while NP(1×10-6 mol/L)decreased intracellular calcium concentration changing amplitude and suppressed cell proliferation. G15 could block the promoting effect of 1×10-10 mol/L NP on the intracel?lular calcium concentration and cell proliferation,but could not block the inhibition of 1×10-6 mol/L NP on the intracellular calcium increase and cell proliferation. Conclusion The results indicate that NP affect rapid calcium signal changes and cell proliferation in non?monotonic dose dependent manner,and its mechanism may be due to the different involvement of GPR30 receptor in different concentrations.

Function and advance of GPR30 in hormone-related neoplasms / 国际肿瘤学杂志

G protein-coupled receptor 30 ( GPR30),a sort of novel functional estrogen transmembrane receptor,extensively participate in the pathological and physiological regulation by mediating the rapid nongenomic effects of the estrogen,which plays an important role in the occurrence and development of the hormone-related malignant neoplasms.