Dysfunction of Shh signaling impaired trophoblast motility and autophagy is involved in poor placentation of recurrent miscarriage

Abstract Background: In early pregnancy, the placenta anchors the conceptus and supports embryonic development and survival. This study aimed to investigate the underlying functions of Shh signaling on recurrent miscarriage, an serious disorder of pregnancy. Methods: Immunofluorescence and immunohistochemistry were used to detect protein expression and its location in placental tissues. Quantitative real-time RT-PCR and Western blot analysis were performed to examine mRNA and protein levels, respectively. Lentiviruses expressing short hairpin RNA were used to knock down the target genes. Cell invasion and migration were performed by with or without Matrigel-coated transwell, respectively. Primary trophoblast migration was performed by villous explant assay. RNA-sequence was used to investigate the genes transcription profile. CCK-8 assay was used to evaluate cell viability. Flow cytometry was used to evaluate cell apoptosis. Results: Our results showed that Shh and Gli2 were mainly located in cytotrophoblasts (CTBs), Ptch was mainly located in syncytiotrophoblasts (STBs), while Smo and Gli3 were expressed in both CTBs and STBs. Compared to the gestational age-matched normal human placenta, the expression of Shh was significantly decreased in recurrent miscarriage. Furthermore, inhibition of Shh signaling impaired motility of JAR cells via regulating the expression of Gli2 and Gli3 to decrease AKT Ser473 phosphorylation, elevate E-cadherin and VEGFA. Intriguingly, inhibition of Shh signaling also enhanced autophagy and autolysosome. Additionally, knockdown BECN1 reversed the effect of Gant61 on motility inhibition. Conclusion: Our results indicated that dysfunction of Shh signaling impaired trophoblast motility, angiogenesis and activated autophagy in villous trophoblast, which would contribute to the pathophysiology of recurrent miscarriage.

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Dysfunction of Shh signaling activates autophagy to inhibit trophoblast motility in recurrent miscarriage

Experimental & Molecular Medicine ◽

10.1038/s12276-020-00530-6 ◽

2021 ◽

Author(s):

Yibin Pan ◽

Lili Yan ◽

Qiaoqiao Chen ◽

Cheng Wei ◽

Yongdong Dai ◽

...

Keyword(s):

Early Pregnancy ◽

Gestational Age ◽

Human Placenta ◽

Recurrent Miscarriage ◽

Shh Signaling ◽

Protein Levels ◽

Shh Pathway ◽

Normal Controls ◽

Jar Cells ◽

Motility Inhibition

AbstractIn early pregnancy, the placenta anchors the conceptus and supports embryonic development and survival. This study aimed to investigate the underlying functions of Shh signaling in recurrent miscarriage (RM), a serious disorder of pregnancy. In the present study, Shh and Gli2 were mainly observed in cytotrophoblasts (CTBs), Ptch was mainly observed in syncytiotrophoblasts (STBs), and Smo and Gli3 were expressed in both CTBs and STBs. Shh signaling was significantly impaired in human placenta tissue from recurrent miscarriage patients compared to that of gestational age-matched normal controls. VEGF-A and CD31 protein levels were also significantly decreased in recurrent miscarriage patients. Furthermore, inhibition of Shh signaling impaired the motility of JAR cells by regulating the expression of Gli2 and Gli3. Intriguingly, inhibition of Shh signaling also triggered autophagy and autolysosome accumulation. Additionally, knockdown of BECN1 reversed Gant61-induced motility inhibition. In conclusion, our results showed that dysfunction of Shh signaling activated autophagy to inhibit trophoblast motility, which suggests the Shh pathway and autophagy as potential targets for RM therapy.

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BCOR Internal Tandem Duplication Expression in Neural Stem Cells Promotes Growth, Invasion, and Expression of PRC2 Targets

International Journal of Molecular Sciences ◽

10.3390/ijms22083913 ◽

2021 ◽

Vol 22 (8) ◽

pp. 3913

Author(s):

Satoshi Nakata ◽

Ming Yuan ◽

Jeffrey A. Rubens ◽

Ulf D. Kahlert ◽

Jarek Maciaczyk ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Tandem Duplication ◽

Target Genes ◽

Cellular Growth ◽

Central Nervous System Tumor ◽

Internal Tandem Duplication ◽

Invasion And Migration ◽

Human Neural Stem Cells ◽

And Migration

Central nervous system tumor with BCL6-corepressor internal tandem duplication (CNS-BCOR ITD) is a malignant entity characterized by recurrent alterations in exon 15 encoding the essential binding domain for the polycomb repressive complex (PRC). In contrast to deletion or truncating mutations seen in other tumors, BCOR expression is upregulated in CNS-BCOR ITD, and a distinct oncogenic mechanism has been suggested. However, the effects of this change on the biology of neuroepithelial cells is poorly understood. In this study, we introduced either wildtype BCOR or BCOR-ITD into human and murine neural stem cells and analyzed them with quantitative RT-PCR and RNA-sequencing, as well as growth, clonogenicity, and invasion assays. In human cells, BCOR-ITD promoted derepression of PRC2-target genes compared to wildtype BCOR. A similar effect was found in clinical specimens from previous studies. However, no growth advantage was seen in the human neural stem cells expressing BCOR-ITD, and long-term models could not be established. In the murine cells, both wildtype BCOR and BCOR-ITD overexpression affected cellular differentiation and histone methylation, but only BCOR-ITD increased cellular growth, invasion, and migration. BCOR-ITD overexpression drives transcriptional changes, possibly due to altered PRC function, and contributes to the oncogenic transformation of neural precursors.

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miR-373-3p Regulates Invasion and Migration Abilities of Trophoblast Cells via Targeted CD44 and Radixin

International Journal of Molecular Sciences ◽

10.3390/ijms22126260 ◽

2021 ◽

Vol 22 (12) ◽

pp. 6260

Author(s):

Hyun-Jung Lee ◽

Seung Mook Lim ◽

Hee Yeon Jang ◽

Young Ran Kim ◽

Joon-Seok Hong ◽

...

Keyword(s):

Preterm Labor ◽

Target Genes ◽

Gene Array ◽

Migration And Invasion ◽

Trophoblast Cells ◽

Invasion And Migration ◽

Mirna Array ◽

Qrt Pcr ◽

Putative Target ◽

And Migration

Preterm labor (PTL) is one of the obstetric complications, and is known to be associated with abnormal maternal inflammatory response and intrauterine inflammation and/or infection. However, the expression of specific miRNAs associated with PTL is not clear. In this study, we performed combination analysis of miRNA array and gene array, and then selected one miRNA (miR-373-3p) and its putative target genes (CD44 and RDX) that exhibited large expression differences in term and PTL placentas with or without inflammation. Using qRT-PCR and luciferase assays, we confirmed that miR-373-3p directly targeted CD44 and RDX. Overexpression of miR-373-3p reduced the migration and invasion of trophoblast cells, while inhibition of miR-373-3p restored the migration and invasion abilities of trophoblast cells. Finally, we validated the expression of miR-373-3p and its target genes in clinical patients’ blood. miR-373-3p was increased in PTL patients’ blood, and was the most expressed in PTL patients’ blood with inflammation. In addition, by targeting the miR-373-3p, CD44 and RDX was decreased in PTL patients’ blood, and their expression were the lowest in PTL patients’ blood with inflammation. Taken together, these findings suggest that miR-373-3p and its target genes can be potential biomarkers for diagnosis of PTL.

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LncRNA MCM3AP-AS1 sponges miR-148a to enhance cell invasion and migration in small cell lung cancer

BMC Cancer ◽

10.1186/s12885-021-08365-8 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Hua Luo ◽

Yukun Zhang ◽

Guangmei Qin ◽

Bing Jiang ◽

Lili Miao

Keyword(s):

Lung Cancer ◽

Small Cell Lung Cancer ◽

Cell Lung Cancer ◽

Cell Invasion ◽

Small Cell ◽

Small Cell Lung ◽

Invasion And Migration ◽

Protein Levels ◽

Underlying Mechanisms ◽

And Migration

Abstract Background MCM3AP-AS1 is a recently characterized lncRNA playing an oncogenic role in several cancers. However, its role in lung cancer remains unknown. Here, we aimed to explore the functions of MCM3AP-AS1 in small cell lung cancer (SCLC) and the possible underlying mechanisms. Methods MCM3AP-AS1 and ROCK1 levels in SCLC patients were analyzed by qPCR. RNA pull-down and luciferase assays were performed to analyze the interaction between MCM3AP-AS1 and miR-148a. ROCK1 mRNA and protein levels were detected by qPCR and Western blot, respectively. Cell invasion and migration were analyzed by Transwell assays. Results MCM3AP-AS1 was upregulated in patients with SCLC, and a high MCM3AP-AS1 level was accompanied by a low survival rate. The binding of MCM3AP-AS1 to miR-148a predicted by bioinformatics analysis was verified by RNA pull-down and luciferase assays. However, MCM3AP-AS1 and miR-148a did not affect each other’s expression. ROCK1 was upregulated in SCLC tissues and positively correlated with MCM3AP-AS1. In SCLC cells, MCM3AP-AS1 overexpression increased ROCK1 and promoted cancer cell invasion and migration, while miR-148a overexpression showed the opposite effects and attenuated the effects of MCM3AP-AS1 overexpression on ROCK1 expression and cell behaviors. Conclusions MCM3AP-AS1 sponges miR-148a, thereby increasing SCLC cell invasion and migration via upregulating ROCK1 expression.

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ERK5 and its role in tumour development

Biochemical Society Transactions ◽

10.1042/bst20110663 ◽

2012 ◽

Vol 40 (1) ◽

pp. 251-256 ◽

Cited By ~ 50

Author(s):

Pamela A. Lochhead ◽

Rebecca Gilley ◽

Simon J. Cook

Keyword(s):

Mitogen Activated Protein Kinase ◽

Invasion And Migration ◽

Extracellular Signal Regulated Kinase ◽

Protein Levels ◽

Extracellular Signal ◽

Mapk Signalling ◽

Mitogen Activated Protein ◽

Tumour Cell Invasion ◽

And Migration

The MEK5 [MAPK (mitogen-activated protein kinase)/ERK (extracellular-signal-regulated kinase) kinase 5]/ERK5 pathway is the least well studied MAPK signalling module. It has been proposed to play a role in the pathology of cancer. In the present paper, we review the role of the MEK5/ERK5 pathway using the ‘hallmarks of cancer’ as a framework and consider how this pathway is deregulated. As well as playing a key role in endothelial cell survival and tubular morphogenesis during tumour neovascularization, ERK5 is also emerging as a regulator of tumour cell invasion and migration. Several oncogenes can stimulate ERK5 activity, and protein levels are increased by a novel amplification at chromosome locus 17p11 and by down-regulation of the microRNAs miR-143 and miR-145. Together, these finding underscore the case for further investigation into understanding the role of ERK5 in cancer.

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miR-17-5p promotes the invasion and migration of colorectal cancer by regulating HSPB2

10.21203/rs.3.rs-63587/v2 ◽

2020 ◽

Author(s):

Wei fang Yu ◽

Jia Wang ◽

Chao Li ◽

Mingda Xuan ◽

Shuangshuang Han ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Proliferation ◽

Lymph Node ◽

Target Genes ◽

Migration And Invasion ◽

Invasion And Migration ◽

Normal Tissues ◽

Node Metastasis ◽

Rescue Experiment ◽

And Migration

Abstract Background: MicroRNA (miRNA) can affect tumor progression by regulating cell proliferation, apoptosis and metastasis. After miRNA microarray chip analysis of colorectal cancer (CRC) tissues and adjacent normal tissues, a significant upregulation of miR-17-5p expression was found in CRC tissues. However, the underlying mechanism of miR-17-5p in CRC is still unclear.Methods: The levels of miR-17-5p in 47 paired CRC and adjacent normal tissue samples were determined by quantitative real-time PCR (qRT-PCR). CCK-8, colony formation, flow cytometry and transwell assays were used to explore the biological effects of miR-17-5p on CRC cells. In addition, the transcriptome sequencing and miRNA target prediction software were employed to identify targets of miR-17-5p. Luciferase reporter detection was used to demonstrate the direct binding of target genes by miR-17-5p. The rescue experiment was conducted to investigate the biological function of target genes and regulatory mechanism of miR-17-5p on target genes.Results: The expression of miR-17-5p was significantly higher in CRC tissues than in adjacent normal tissues. In CRC group, the expression of miR-17-5p in cancer tissues with lymph node metastasis was higher compared with those without lymph node metastasis. Overexpression of miR-17-5p inhibited CRC cell apoptosis, as well as promoting proliferation, migration and invasion. We hypothesized that HSPB2 might be a target gene of miR-17-5p and validated for the first time that miR-17-5p binds directly to the 3’-UTR of HSPB2. In the rescue experiment, the tumor suppressive effect of HSPB2 was detected and miR-17-5p could promote cell proliferation, migration and invasion by targeting HSPB2.Conclusion: MiR-17-5p promotes invasion and migration by inhibiting HSPB2 in CRC, thereby implicating its potential as a novel diagnostic biomarker and therapeutic target for CRC.

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Id-2 regulates critical aspects of human cytotrophoblast differentiation, invasion and migration

Development ◽

10.1242/dev.127.3.549 ◽

2000 ◽

Vol 127 (3) ◽

pp. 549-558 ◽

Cited By ~ 1

Author(s):

M.J. Janatpour ◽

M.T. McMaster ◽

O. Genbacev ◽

Y. Zhou ◽

J. Dong ◽

...

Keyword(s):

Dna Binding ◽

Adenovirus Vector ◽

Bhlh Transcription Factor ◽

Placental Development ◽

Invasion And Migration ◽

Protein Levels ◽

Hypoxic Conditions ◽

Bhlh Factors ◽

Cytotrophoblast Cells ◽

And Migration

During early human placental development, the conceptus attaches itself to the uterus through cytotrophoblast invasion. Invasive cytotrophoblast cells differentiate from precursor villous cytotrophoblasts, but the essential regulating factors in this process are unknown. Basic helix-loop-helix (bHLH) transcription factor dimers are essential regulators of mouse trophoblast development. We therefore examined the importance of this family of factors in the human placenta. In many cell lineages, bHLH factors are sequestered by members of the Id family, HLH proteins that lack the basic DNA binding domain (Inhibitor of DNA binding proteins (Id-1 to Id-4)). During differentiation of some tissues, Id expression declines, allowing bHLH factors to dimerize, bind DNA and trans-activate lineage-specific genes. To begin to study the role of bHLH transcription factors in human placental development, we first characterized Id expression in cytotrophoblast cells. The cells expressed Id-3 constitutively; Id-2 was downregulated, at the mRNA and protein levels, as the cells differentiated in culture and in situ, respectively. In cases when cytotrophoblast differentiation was compromised (in placentas from women with preeclampsia, or in cells grown under hypoxic conditions in culture), Id-2 expression was maintained. To assess the functional relevance of these correlations, we used an adenovirus vector to maintain Id-2 protein expression in cultured cytotrophoblasts. Compared to control (lacZ-expressing) cells, cytotrophoblasts transduced to constitutively express Id-2 retained characteristics of undifferentiated cells: (alpha)1 integrin expression was low and cyclin B expression was retained. Furthermore, invasion through Matrigel was partially inhibited and migration was strikingly enhanced in Id-2-expressing cells. These results suggest that Id-2 and the bHLH factors that it partners play important roles in human cytotrophoblast development.

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Annexin A2 Coordinates STAT3 to Regulate the Invasion and Migration of Colorectal Cancer CellsIn Vitro

Gastroenterology Research and Practice ◽

10.1155/2016/3521453 ◽

2016 ◽

Vol 2016 ◽

pp. 1-10 ◽

Cited By ~ 11

Author(s):

Dianhui Xiu ◽

Lin Liu ◽

Fengli Qiao ◽

Haishan Yang ◽

Lu Cui ◽

...

Keyword(s):

Colorectal Cancer ◽

Annexin A2 ◽

Migration And Invasion ◽

Invasion And Migration ◽

Protein Levels ◽

And Migration

The present study aimed to reveal the expression of STAT3 and Anxa 2 in CRC specimens and to investigate the effects of STAT3 and Anxa 2 signaling on the proliferation, invasion, and migration in CRC Caco-2 cells. Results demonstrated that both Anxa 2 and STAT3 were highly expressed in CRC specimens in both mRNA and protein levels, with or without phosphorylation (Tyrosine 23 in Anxa 2 and Tyrosine 705 in STAT3). And the upregulated Anxa 2 promoted the phosphorylation of STAT3 (Tyrosine 705) in CRC Caco-2 cells. The upregulated Anxa 2 promoted the proliferation, migration, and invasion of Caco-2 cells in vitro. Moreover, the STAT3 knockdown also repressed the proliferation, migration, and invasion of Caco-2 cells. In conclusion, the overexpressed Annexin A2 regulated the proliferation, invasion, and migration in CRC cells in an association with STAT3.

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Knockdown of TPPP3 to inhibit cell proliferation and invasion in human colorectal cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.15_suppl.e23006 ◽

2017 ◽

Vol 35 (15_suppl) ◽

pp. e23006-e23006 ◽

Cited By ~ 2

Author(s):

Yintao Li ◽

Jinming Yu

Keyword(s):

Colorectal Cancer ◽

Cell Proliferation ◽

Protein Family ◽

Migration And Invasion ◽

Clinicopathologic Characteristics ◽

Invasion And Migration ◽

Protein Levels ◽

Angiogenesis Assay ◽

And Migration

e23006 Background: Tubulin Polymerization Promoting Protein Family Member 3, TPPP3, a member of the TPPP protein family, has been reported to play important roles in initiation and progression of human cancers, such as lung cancer. However, the expression and underlying function of TPPP3 in colorectal cancer (CRC) have not yet been fully clarified. Methods: In this study, the mRNA and protein levels of TPPP3 in 96 clinical CRC specimens were determined by RT-PCR and immunohistochemistry. The relation between TPPP3 expression and clinicopathologic characteristics and overall survival (OS) were evaluated. TPPP3 was stably knockdowned by shRNA. In addition, CCK-8、Colony formation、Flow cytometric、Transwell and Angiogenesis assay were to examine the biological function of TPPP3 in CRC cells in vitro. Results: We show that TPPP3 was significantly increased in CRC tissues and associated with aggressive factors and poor patient survival. Further experiments showed that knockdown of TPPP3 inhibited cell proliferation, migration and invasion and induced cell apoptosis in vitro. In addition, TPPP3 silencing resulted in a decrease of angiogenesis and S phase fraction. And TPPP3 significantly affected the invasion and migration of CRC cells via the expression of MMP-9, Rac-1 and E-cadherin. Conclusions: Our results suggested that TPPP3 played an important role in CRC progress and might serve as novel therapeutic targets for CRC treatment.

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