Secukinumab plays a synergistic role with starvation therapy in promoting autophagic cell death of hepatocellular carcinoma via inhibiting IL-17A-increased BCL2 level.

It is known that IL-17A inhibits autophagy of hepatocellular carcinoma (HCC) cells, thus contributing to the carcinogenesis of HCC. Starvation therapy can promote the autophagic death of HCC cells by blocking the nutrition supply. The purpose of this study was to explore whether the pharmacological antagonist of IL-17A, secukinumab, and starvation therapy have a synergistic effect on the autophagic cell death of HCC. Here, it could be observed that compared with serum-free condition, the combination of secukinumab and serum-free status better promoted autophagy (observed by LC3 conversion rate, p62 protein expression and the formation of autophagosomes), and more significantly inhibited the survival and function (observed by Trypan blue staining, CCK-8, Transwell, and scratch assays) in HCC HepG2 cells. Moreover, secukinumab significantly decreased BCL2 protein expression under serum-normal and serum-free conditions. However, both the addition of recombinant IL-17A and overexpression of BCL2 blocked the regulation of secukinumab on the survival and autophagy in HepG2 cells. Nude mice experiments demonstrated that compared to the lenvatinib-alone group, the combination group of lenvatinib and secukinumab better inhibited the in vivo tumorigenesis of HepG2 cells and enhanced autophagy in xenotumor tissues. Furthermore, secukinumab significantly decreased BCL2 protein expression in xenotumor tissues without or with lenvatinib application. In conclusion, the antagonism of IL-17A with secukinumab, due to the upregulation on BCL2-related autophagic cell death, can cooperate with starvation therapy in inhibiting HCC carcinogenesis. Our data suggested that secukinumab can become an effective adjuvant for the treatment of HCC.

CD142 promotes trophoblast cell migration by inhibiting BCL2-related autophagic degradation of IL-8.

The maintenance of migration of trophoblast cells is beneficial to pregnancy, and its weakening can lead to preeclampsia (PE). CD142 is considered as a classical motility-promoting factor. Our research aimed to explore the role of CD142 in trophoblast cell migration and potential mechanism. Through fluorescence-activated cell sorting (FACS) and gene transduction assays, CD142 expression levels of mouse trophoblast cell lines were upregulated and downregulated respectively. Then, the migratory level was detected through Transwell assays in different groups of trophoblast cells. The corresponding chemokines were screened by ELISA in different sorted trophoblast cells. Based on gene overexpression and knockdown assays, the production mode of identified valuable chemokine was analyzed by detecting gene and protein expression in trophoblast cells. Finally, the contribution of autophagy response to specific chemokine regulated by CD142 was explored by combining different groups of cells and autophagy regulators. Our results showed that both CD142 positive sorting and CD142 overexpression promoted the migratory ability of trophoblast cells, and trophoblast cells with the highest level of CD142 had the strongest migratory ability. In addition, CD142+ cells contained the highest level of IL-8. Consistently, CD142 overexpression promoted IL-8 protein expression in trophoblast cells while CD142 silencing was contrary. However, both CD142 overexpression and CD142 silencing did not affect IL-8 mRNA expression. Moreover, both CD142+ and CD142-overexpressed cells showed higher BCL2 protein expression and poorer autophagic activity. Importantly, autophagy activation with TAT-Beclin1 recovered the increased IL-8 protein expression in CD142+ cells. Obviously, the migratory ability of CD142+ cells inhibited by TAT-Beclin1 was recovered by the addition of IL-8 recombinant factor. In conclusion, CD142 inhibits the degradation of IL-8 through the inhibition of BCL2-Beclin1-autophagy signal transduction, thereby promoting the migration of trophoblast cells.

The relationship between hsa_circ_0051326 and HLA-G expression in the blood of patients with pre-eclampsia.

OBJECTIVES: To investigate the relationship between hsa_circ_0051326 and HLA-G expression in the blood of patients with pre-eclampsia. MATERIAL AND METHODS: Real-time PCR (qRT-PCR) was used to detect the hsa_circ_0051326 expression level. Enzyme-linked immunosorbent assay (ELISA), qRT-PCR, and western blotting were used to detect HLA-G expression in blood in 50 patients with pre-eclampsia and 50 normal pregnant women. RESULTS: HLA-G protein expression level was decreased significantly in the blood of patients with pre-eclampsia. hsa_circ_0051326 and HLA-G mRNA in blood were decreased significantly in the pre-eclampsia patients compared with normal pregnant women. There was a positive correlation between the expression of serum hsa_circ_0051326 with HLA-G mRNA. CONCLUSIONS: Serum hsa_circ_0051326 was a new diagnostic biomarker for pre-eclampsia with equivalent efficiency of HLA-G. Maternal serum hsa_circ_0051326 level may pre-diagnose potentially pre-eclampsia before delivery.

Inhibition of the cell migration, invasion and chemoresistance of colorectal cancer cells through targeting KLF3 by miR-365a-3p.

Background: Metastasis and chemoresistance limit treatment efficacy of colorectal cancer (CRC) patients. MicroRNAs (miRNAs) have been believed to be candidate biomarkers for tumor cell proliferation, metastasis and chemoresistance, but the related molecular mechanisms are not clear for prognosis prediction. Aims: We aimed to investigate the role of miR-365a-3p in metastasis and chemoresistance of CRC. Methods: The expression levels of miR-365a-3p in clinical CRC tissues were analyzed. The effects of miR-365a-3p expression levels on tumor chemoresistance, invasion and migration were also determined. A dual luciferase reporter gene assay was used to determine the effect of miR-365a-3p on its target gene, Kruppel-like factor 3 (KLF3), and the effect of the miR-365a-3p/KLF3 axis on CRC cell chemoresistance, migration and invasion was further investigated. Results: In patients with CRC with lymph node or distant organ metastasis or in CRC cell lines, the expression levels of miR-365a-3p were significantly downregulated. In addition, the findings of Transwell assays demonstrated that miR-365a-3p significantly suppressed CRC cell migration and invasion. The dual luciferase reporter gene assay results suggested that miR-365a-3p may play an important role in the regulation of migration, invasion and chemoresistance in CRC cells. Conclusions: The findings of present study provided evidence to suggest that miR-365a-3p may be a potential tumor suppressor gene in CRC and may inhibit the migration, invasion and chemoresistance of CRC cells. These results suggested that targeting miR-365a-3p/KLF3 axis may represent a potential therapeutic intervention for metastatic disease in patients with CRC.

Significance of highly phosphorylated insulin-like growth factor binding protein-1 and cervical length for prediction of preterm delivery in twin pregnancies.

BACKGROUND: A twin pregnancy can carry greater risks than singleton pregnancies. About 60 in 100 twin pregnancies result in spontaneous birth before 37 wk, which is associated with several complications in the premature babies. Clinical detection of biomarkers may help to predict the possibility of premature birth so that corresponding interventions can be given to the pregnant women in a timely manner, in order to reduce the risk of preterm birth and improve the outcomes of the newborn infants. AIM: To explore the clinical value of transvaginal ultrasound measurement of cervical length combined with insulin-like growth factor binding protein-1 (IGFBP-1) hyperphosphorylation in cervical secretions as predictors of preterm delivery in twin pregnancies. METHODS: A total of 254 pregnant women with twin pregnancies, who were admitted to Hainan General Hospital and underwent maternity examination, were selected as the study subjects from January 2015 to December 2018. All participants received transvaginal ultrasound measurement of cervical length and phosphorylated IGFBP-1 (phIGFBP-1) test between 24 and 34 wk gestation. The pregnancy outcomes were analyzed. RESULTS: Of the women with a positive phIGFBP-1 test result, preterm birth rate was higher in those with a cervical length ≤ 25 mm than those with a cervical length > 25 mm (all P < 0.05). Similarly, in women with a negative phIGFBP-1 test result, preterm birth rate was higher in those with a cervical length ≤ 25 mm than those with a cervical length > 25 mm (all P < 0.05). The sensitivity, specificity, and positive and negative predictive values of the phIGFBP-1 test combined with the cervical length test were 95.71%, 91.21%, 95.12% and 92.22%, respectively, for the prediction of preterm birth. CONCLUSION: Cervical length combined with phIGFBP-1 tests is of value for the prediction of outcomes of preterm delivery in twin pregnancies.

MicroRNA-490-3p inhibits migration and chemoresistance of colorectal cancer cells via targeting TNKS2.

OBJECTIVE: Colorectal cancer is one of the most common malignancy in the world. The oncogenesis of colorectal cancer is still not fully elucidated. It was reported that microRNA-490-3p (miR-490-3p) was closely related to the regulation of cancers. However, if miR-490-3p could also affect colorectal cancer and the specific mechanism remains unclear. METHODS: qRT-PCR was conducted to examine the expression of miR-490-3p. DIANA, miRDB, and TargetScan databases were used to identify target genes. LOVO and SW480 cells were transfected by miR-490-3p mimics and inhibitors. Transwell assay was used to measure cell invasion and migration. Cisplatin and fluorouracil were administered to investigate chemotherapy resistance. Western blot was used to measure TNKS2 protein expression. Binding sites were verified using the double luciferase assay. RESULTS: miR-490-3p expression was low in the colorectal cancer cells. The level of miR-490-3p was negatively correlated with cell migration and invasion of cancer cells. miR-490-3p could bind to TNKS2 mRNA 3'UTR directly. miR-490-3p can suppress cell viability and resistance to chemotherapy in colorectal cancer cells through targeting TNKS2. CONCLUSIONS: miR-490-3p could affect colorectal cancer by targeting TNKS2. This study may provide a potential therapeutic target for colorectal cancer.

MicroRNA­524­5p regulates the proliferation and invasion of HTR­8/SVneo trophoblasts by targeting NUMB in the Notch signaling pathway.

Preeclampsia is a pregnancy disorder that is primarily associated with maternal and neonatal or fetal morbidity and mortality. The discovery of dysregulated microRNAs (miRs) and their roles in preeclampsia has provided new insight into the mechanisms involved in pregnancy­related disorders. In the present study, quantitative PCR demonstrated that the expression levels of miR­524­5p were lower in patients with preeclampsia than those in normal pregnant women. Cell Counting Kit­8 and Transwell assays indicated that overexpression of miR­524­5p promoted the proliferation and invasion of HTR­8/SVneo cells, whereas inhibition of miR­524­5p suppressed HTR­8/SVneo cell proliferation and invasion. Furthermore, NUMB endocytic adaptor protein (NUMB), a negative regulator of the Notch signaling pathway and a target gene of miR­524­5p, limited the effects of miR­524­5p on HTR­8/SVneo cell invasion and migration. The present study demonstrated that miR­524­5p regulated the proliferation and invasion of HTR­8/SVneo cells at least partly by targeting NUMB to regulate the Notch signaling pathway.

Vagus nerve stimulation ameliorates L-NAME-induced preeclampsia-like symptoms in rats through inhibition of the inflammatory response.

BACKGROUND: Preeclampsia is characterized by an excessive inflammatory response. Recent studies have shown that vagus nerve stimulation (VNS) has anti-inflammatory properties in vivo. This study aims to investigate whether VNS is safe for use during pregnancy and to explore the therapeutic potential and underlying mechanisms of VNS in PE. METHODS: Pregnant Sprague-Dawley rats were randomly chosen to receive N-nitro-L-arginine methyl ester (L-NAME)-containing water (preeclampsia-like mouse model) or saline (normal pregnancy control) daily at gestational days 14.5-20.5. VNS and the α7nAChR antagonist methyllycaconitine citrate (MLA, 1 mg/kg/d) were given daily at the same time. RESULTS: VNS decreased the high systolic blood pressure and urinary protein observed in the PE rats. In addition, VNS mitigated abnormal pregnancy outcomes. Moreover, VNS alleviated the inflammatory response by decreasing the levels of inflammatory cytokines. VNS significantly increased the expression of α7nAChR and attenuated the activation of NF-κB p65 in the placenta. DISCUSSION: Our findings indicate that maternal VNS treatment is safe during pregnancy and has a protective effect in a pregnant rat model of preeclampsia induced by L-NAME.

Placental expression of AChE, α7nAChR and NF-κB in patients with preeclampsia.

OBJECTIVES: This study aimed to investigate placental expression of AChE, α7nAChR and NF-κB in patients with preeclampsia and discuss about its clinical significance. MATERIAL AND METHODS: mRNA expression levels of acetylcholine (AChE), alpha-7 nicotinic acetylcholine receptor (α7nAChR) and nuclear factor-kB (NF-κB) in placenta were detected by qRT-PCR, and protein levels were determined by immunohis-tological analysis and Western Blot in 35 women with preeclampsia (including 20 cases of mild preeclampsia and 15 cases of severe preeclampsia) and 30 cases in control group, respectively. RESULTS: The expression of AChE mRNA and protein in placenta increased significantly in patients with preeclampsia compared with the control group (p < 0.01). It was lower in patients with severe preeclampsia than in patients with mild preeclampsia (p < 0.05). The expression of α7nAChR mRNA and protein in placenta decreased significantly in patients with preeclampsia compared with the control group (p < 0.01). However, the expression of α7nAChR mRNA and protein in patients with severe preeclampsia was higher than that in patients with mild preeclampsia, without significant difference(p > 0.05). The expression of NF-κB protein in placenta decreased significantly in patients with preeclampsia compared with the control group(p < 0.01). It was higher in patients with severe preeclampsia than in patients with mild preeclampsia (p < 0.05), but there was no significant difference between preeclampsia group and control group in the expression of NF-κB mRNA in placenta (p > 0.05). The results of Western blotting assay were consistent with those of immunohistochemistry. CONCLUSIONS: Abnormal expression of AChE, α7nAChR and NF-κB in placenta may be associated with preeclampsia. Cho-linergic anti-inflammatory pathway may play an important role in the pathogenesis of preeclampsia.