CCND2 and miR-206 as potential biomarkers in the clinical diagnosis of thyroid carcinoma by fine-needle aspiration cytology.

BACKGROUND: To investigate the relationship between cyclin D2 (CCND2) and miR-206 expression in fine-needle aspiration cytology of thyroid carcinoma. METHODS: A total of 65 patients with thyroid carcinoma were selected as the subjects and 65 patients with benign thyroid nodules were in control group. The fine-needle aspiration cytology of thyroid nodules was performed. CCND2 and miR-206 levels were detected by PCR. RESULTS: Compared with the patients with benign thyroid nodules, the expression level of miR-206 in fine-needle aspiration cytology of thyroid cancer patients decreased significantly and the expression level of CCND2 increased significantly. CCND2 and miR-206 expression was negatively correlated in thyroid cancer tissues. Area under curve (AUC) of miR-206 level in the diagnosis of thyroid cancer was 0.889, and the sensitivity and specificity were 92.3% and 81.5%, respectively. AUC of CCND2 level in the diagnosis of thyroid cancer was 0.837, and the sensitivity and specificity were 67.7% and 89.2%, respectively. The AUC of combined detection of CCND2 and miR-206 in the diagnosis of thyroid cancer was 0.959, and the sensitivity and specificity were 93.8% and 87.7%, respectively. The levels of miR-206 and CCND2 were significantly correlated with TNM staging and lymph node metastasis. CONCLUSIONS: miR-206 and CCND2 may become new biomarkers for clinical diagnosis of thyroid cancer based on the fine-needle aspiration cytology of thyroid nodules.

Substance P Promotes the Progression of Endometrial Adenocarcinoma.

OBJECTIVES: It has been demonstrated that substance P (SP) promotes while neurokinin-1 receptor (NK-1R) antagonist inhibits the proliferation of several human cancer cells. Currently, it is still unknown whether such actions exist in human endometrial carcinoma. This study aimed to explore the role of SP/NK-1R signaling in the progression of endometrial adenocarcinoma. MATERIALS AND METHODS: The expression levels of SP and NK-1R in endometrial adenocarcinoma tissues and Ishikawa cell line were detected by real-time quantitative PCR and Western blot analysis. The effects of SP on Ishikawa cells proliferation and invasion were analyzed using MTT assay and transwell matrigel invasion assay, respectively. The expression levels of matrix metalloproteinase 9 (MMP-9) and vascular endothelial growth factor C (VEGF-C) in Ishikawa cells after administration of SP were detected by real-time quantitative RCR and Western blot analysis. RESULTS: The expression levels of SP and NK-1R were significantly higher in endometrial adenocarcinoma tissues and Ishikawa cells than in normal endometrium. Substance P significantly enhanced the proliferation and invasion of Ishikawa cells. In addition, SP induced the expression of MMP-9 and VEGF-C in Ishikawa cells, whereas NK-1R antagonist inhibited these effects. CONCLUSIONS: Substance P plays an important role in the development of endometrial carcinoma by inducing the expression of MMP-9 and VEGF-C and promoting cancer cell proliferation and metastasis, which can be blocked by NK-1R antagonist.

Substance P is overexpressed in cervical squamous cell carcinoma and promoted proliferation and invasion of cervical cancer cells in vitro.

This study aimed to investigate the expression and function of substance P in cervical squamous cell carcinoma. Cancer tissues and adjacent tissues of 20 patients with cervical squamous cell carcinoma in our hospital were collected. The expression of substance P was detected by immunohistochemistry and Western blot analysis. Cervical squamous cell carcinoma line SiHa was treated with different concentrations of substance P. The proliferation of SiHa cells was detected by EdU assay, and the invasion ability of SiHa cells was detected by transwell assay. The phosphorylation of ERK1/2 and the expression of MMP9 were detected by Western blot analysis. The results showed that substance P was expressed in the cytoplasm and some cell membranes of cervical squamous cell carcinoma cells. The expression of substance P in cervical cancer tissues was significantly higher than that in the adjacent tissues. Compared with the control group, substance P significantly promoted the proliferation and invasion of SiHa cells in a concentration dependent manner and activated the phosphorylation of ERK1/2 and upregulated the expression of MMP9 in SiHa cells. In conclusion, substance P is highly expressed in cervical squamous cell carcinoma and can promote cervical cancer cell proliferation and invasion. The mechanism is related to the activation of ERK1/2 pathway to upregulate MMP9.

Correlation between thyroid function indices and urine iodine levels in patients with nodular goiter.

OBJECTIVE: To retrospectively analyze the iodine nutritional status in patients with nodular goiter (NG) and investigate a possible association between urinary iodine levels and thyroid function indices. METHODS: A total of 173 patients diagnosed with nodular goiter in the Fourth Hospital of Hebei Medical University from January 2019 to May 2021 were selected as the NG group, and 172 healthy individuals without thyroid diseases were selected after a physical examination as a control group. The data of all the participants were retrospectively assessed to explore the association between urinary iodine levels and thyroid function indices. The content of urinary iodine in the two groups was compared, and the correlation of urinary iodine levels with thyroid stimulating hormone (TSH), free triiodothyronine (FT3), and free thyroxine (FT4) in the NG group was evaluated. RESULTS: The level of urinary iodine in the NG group was 163.97 ± 113.75 µg/L, which was higher than 121.47 ± 53.75 µg/L in the control group (P < 0.05). The iodine excess rate in females was higher than that in males (P < 0.05). The results of Pearson correlation analysis showed that the amount of urinary iodine in patients with hyperthyroidism with different urinary iodine statuses was negatively correlated with the level of TSH and positively correlated with levels of FT3 and FT4. CONCLUSION: There is a significant association between urinary iodine levels and thyroid hormone levels in NG patients. Therefore, regular monitoring of urinary iodine levels is essential for the appropriate use of iodine supplementation.

Neurokinin-1 receptor is highly expressed in cervical cancer and its antagonist induces cervical cancer cell apoptosis.

Neurokinin-1 receptor (NK1R) belongs to tachykinin receptor family. Recent studies have suggested that NK1R was upregulated in cancer tissues including breast cancer, glioma and melanoma. Furthermore, NK1R antagonists have been employed to exert anti-tumor effect and promote cancer cell apoptosis. However, the role of NK1R in cervical cancer remains largely unknown. In this study, we aimed to detect the expression of NK1R in cervical cancer and evaluate the anti-tumor effects of NK1R antagonist on cervical cancer cells. We found that NK1R was highly expressed in cervical cancer tissues than in adjacent normal cervical tissues. Furthermore, by using NK1R antagonist we demonstrated that NK1R antagonist inhibited the viability and induced the apoptosis of cervical cancer cells in a dose-dependent manner, and the mechanism may be related to the inhibition of ERK activation and the regulation of apoptosis proteins Bcl-2 and BAX. In conclusion, these findings suggest that NK1R plays an oncogenic role in cervical cancer and is a promising target for cervical cancer therapy.

WIF1 was downregulated in cervical cancer due to promoter methylation.

Wnt inhibitory factor 1 (WIF1) is frequently downregulated in a variety of cancer due to promoter methylation. However, the methylation status of the WIF1 promoter in cervical cancer remains unclear. This study aimed to elucidate the mechanism by which WIF1 promoter methylation contributes to cervical cancer development. The expression of WIF1 in cervical cancer tissues was examined by immunohistochemistry. The methylation status of the WIF1 promoter in cervical cancer cells was detected by methylation specific PCR. WIF1 mRNA levels and protein levels were detected by PCR and Western blot analysis. We found that WIF1 expression was low in cervical cancer tissues compared to adjacent normal cervical tissues. The WIF1 promoter was methylated in the cervical cancer SiHa cell line but not in the normal cervical epithelial cell line Ect1. Correspondingly, WIF1 mRNA levels and protein levels were significantly lower in SiHa cells than in Ect1 cells. Treatment with 5-aza-2-deoxycytidine (AZA) led to the upregulation of WIF1 mRNA and protein levels in SiHa cells, but the effects were abrogated by treatment with WIF1 siRNA. In addition, AZA treatment induced apoptosis and inhibited the invasion of SiHa cells, and the effects were abrogated by WIF1 siRNA. The protein levels of survivin, c-myc and cyclinD1 were significantly lower in SiHa cells treated with AZA, but their levels were upregulated after treatment with WIF1 siRNA. In conclusion, the methylation of the WIF1 promoter leads to the downregulation of WIF1 and the activation of Wnt/ß-catenin signaling in cervical cancer cells. WIF1 is a tumor suppressor that is inactivated in cervical cancer.

OP9-DL1 cell co-culture enhances anti-tumour immunity of mouse bone marrow-derived dendritic cells.

DCs (dendritic cells) are the strongest professional APCs (antigen-presenting cells) to initiate immune responses against pathogens, but they are usually incompetent in initiating efficient immune responses in the progress of solid tumours. We have shown that Notch signalling plays a pivotal role in DC-dependent anti-tumour immunity. Compared with the control DCs, OP9-DL1 (Delta-like1) cell co-cultured DCs gained increased tumour suppression activity when inoculated together with tumour cells. This was probably due to the activation of Notch signalling in DCs enhancing their ability to evoke anti-tumour immune responses in solid tumours. Indeed, the OP9-DL1 cell co-cultured DCs expressed higher levels of MHC I, MHC II, CXCR4 (CXC chemokine receptor 4), CCR7 (CC chemokine receptor 7), IL-6 (interleukin 6), IL-12 and TNFα (tumour necrosis factor α), and a lower level of IL-10 than control DCs, resulting in more efficient DC migration and T-cell activation in vivo and in vitro. T-cells stimulated by OP9-DL1 cells co-cultured DCs more efficiently; and were cytotoxic against tumour cells, in contrast with control DCs. These results indicated that up-regulation of Notch signalling in DCs by co-culturing with OP9-DL1 cells enhances DC-dependent anti-tumour immune reactions, making the Notch signalling pathway a target for the establishment of the DC-based anti-tumour immunotherapies.

Substance P immunoreactive nerve fibres are related to gastric cancer differentiation status and could promote proliferation and migration of gastric cancer cells.

Tachykinins such as SP (substance P) may be involved in the progression of gastric adenocarcinoma through binding to NK-1 receptor. However, the existence and relationship between SP and gastric cancer progression and differentiation remained unknown. We have studied the NK-1 receptor in human gastric cancer tissue and MKN45 cell line and found SP-containing nerve fibres in human gastric cancer and found that the amounts of SP-positive nerves were related to gastric cancer differentiation. SP could promote proliferation, adhesion, migration and invasion of MKN45 cells in vitro. In addition, the intracellular calcium level of MKN45 cells was elevated after SP stimulation, and administration of CRACs (calcium release-activated calcium channels) inhibitor SKF-96365 could partially abolish these effects induced by SP. These results demonstrated that NK-1 receptor and SP-containing nerves existed in human gastric cancer; SP positive nerves may play an important role in human gastric cancer progression, and calcium is critically significant among SP-induced biological effects.