Anosmin-1 contributes to brain tumor malignancy through integrin signal pathways

Choy, Catherine T,Kim, Haseong,Lee, Ji-Young,Williams, David M,Palethorpe, David,Fellows, Greg,Wright, Alan J,Laing, Ken,Bridges, Leslie R,Howe, Franklyn A,Kim, Soo-Hyun Bioscientifica Ltd 2014 Endocrine-related cancer Vol.21 No.1

<P>Anosmin-1, encoded by the <I>KAL1</I> gene, is an extracellular matrix (ECM)-associated protein which plays essential roles in the establishment of olfactory and GNRH neurons during early brain development. Loss-of-function mutations of <I>KAL1</I> results in Kallmann syndrome with delayed puberty and anosmia. There is, however, little comprehension of its role in the developed brain. As reactivation of developmental signal pathways often takes part in tumorigenesis, we investigated if anosmin-1-mediated cellular mechanisms associated with brain tumors. Our meta-analysis of gene expression profiles of patients' samples and public microarray datasets indicated that <I>KAL1</I> mRNA was significantly upregulated in high-grade primary brain tumors compared with the normal brain and low-grade tumors. The tumor-promoting capacity of anosmin-1 was demonstrated in the glioblastoma cell lines, where anosmin-1 enhanced cell motility and proliferation. Notably, anosmin-1 formed a part of active β1 integrin complex, inducing downstream signaling pathways. ShRNA-mediated knockdown of anosmin-1 attenuated motility and growth of tumor cells and induced apoptosis. Anosmin-1 may also enhance the invasion of tumor cells within the ECM by modulating cell adhesion and activating extracellular proteases. In a mouse xenograft model, anosmin-1-expressing tumors grew faster, indicating the role of anosmin-1 in tumor microenvironment <I>in vivo</I>. Combined, these data suggest that anosmin-1 can facilitate tumor cell proliferation, migration, invasion, and survival. Therefore, although the normal function of anosmin-1 is required in the proper development of GNRH neurons, overexpression of anosmin-1 in the developed brain may be an underlying mechanism for some brain tumors.</P>

Effect of pioglitazone on serum concentrations of osteoprotegerin in patients with type 2 diabetes mellitus

Park, Jong Suk,Cho, Min Ho,Nam, Ji Sun,Yoo, Jeong Seon,Ahn, Chul Woo,Cha, Bong Soo,Kim, Kyung Rae,Lee, Hyun Chul BioScientifica 2011 European journal of endocrinology Vol.164 No.1

<P><B>Objective</B></P><P>Osteoprotegerin (OPG) acts as an important regulatory molecule in atherosclerosis. Recent studies report that thiazolidinediones could affect OPG expression. We investigated the relationship between OPG and inflammatory cytokines and the effects of pioglitazone (a PPARγ (PPARG) agonist) versus metformin on serum OPG levels in type 2 diabetic patients.</P><P><B>Design and methods</B></P><P>Sixty-seven type 2 diabetic patients were included in this study. They were assigned to pioglitazone (15 mg/day, <I>n</I>=34) or metformin (1000 mg/day, <I>n</I>=33) during 24 weeks. Various anthropometric and metabolic parameters, OPG, interleukin 6 (IL6), C-reactive protein (CRP), adiponectin, and homeostasis model assessment of insulin resistance (HOMA-IR), were measured at baseline and at 6 months of treatment.</P><P><B>Results</B></P><P>Serum OPG levels correlated significantly with fasting plasma glucose (FPG), HbAlc, HOMA-IR, IL6, and CRP, and inversely correlated with adiponectin after adjusting for age (<I>P</I><0.05). Multiple regression analysis showed that FPG, HbAlc, and adioponectin were independently correlated with OPG level. After 6 months of treatment, the reduction in FPG and HbAlc levels was similar between the two groups. Pioglitazone treatment significantly increased body mass index (<I>P</I><0.05) and waist circumference (<I>P</I><0.05) and decreased triglycerides (<I>P</I><0.05) and HOMA-IR (<I>P</I><0.01). The adiponectin concentration was increased (<I>P</I><0.05), and OPG and CRP levels were decreased in the pioglitazone group (<I>P</I><0.05), but were unchanged in the metformin group. The changes in serum OPG in the pioglitazone group showed significant correlation with changes in FPG, HbAlc, and adiponectin.</P><P><B>Conclusions</B></P><P>In type 2 diabetic patients, pioglitazone decreases OPG levels, and this decrease in OPG levels might be associated with the increase in adiponectin.</P>

Distinct roles of <i>ROCK1</i> and <i>ROCK2</i> during development of porcine preimplantation embryos

Zhang, Jin Yu,Dong, Huan Sheng,Oqani, Reza K,Lin, Tao,Kang, Jung Won,Jin, Dong Il BioScientifica 2014 Reproduction Vol.148 No.1

<P>Cell-to-cell contact mediated by cell adhesion is fundamental to the compaction process that ensures blastocyst quality during embryonic development. In this study, we first showed that Rho-associated coiled-coil protein kinases (ROCK1 and ROCK2) were expressed both in porcine oocytes and IVF preimplantation embryos, playing different roles in oocytes maturation and embryo development. The amount of mRNA encoding ROCK1 and the protein concentration clearly increased between the eight-cell and morula stages, but decreased significantly when blastocysts were formed. Conversely, ROCK2 was more abundant in the blastocyst compared with other embryonic stages. Moreover, immunostaining showed that ROCK1 protein distribution changed as the embryo progressed through cleavage and compaction to the morula stage. Initially, the protein was predominantly associated with the plasma membrane but later became cytoplasmic. By contrast, ROCK2 protein was localized in both the cytoplasm and the spindle rotation region during oocyte meiosis, but in the cytoplasm and nucleus as the embryo developed. In addition, ROCK2 was present in the trophectoderm cells of the blastocyst. Treatment with 15 μM Y27632, a specific inhibitor of ROCKs, completely blocked further development of early four-cell stage embryos. Moreover, we did not detect the expression of <I>ROCK1</I> but did detect <I>ROCK2</I> expression in blastocysts. Moreover, lysophosphatidic acid an activator of ROCKs significantly improved the rates of blastocyst formation. These data demonstrate that ROCKs are required for embryo development to the blastocyst stage. Together, our results indicate that ROCK1 and ROCK2 may exert different biological functions during the regulation of compaction and in ensuring development of porcine preimplantation embryos to the blastocyst stage.</P>

Lowered cutoff of lymph node fine-needle aspiration thyroglobulin in thyroid cancer patients with serum anti-thyroglobulin antibody

Jo, Kwanhoon,Kim, Min-Hee,Lim, Yejee,Jung, So-Lyung,Bae, Ja-Seong,Jung, Chan-Kwon,Kang, Moo-Il,Cha, Bong-Yun,Lim, Dong-Jun BioScientifica Ltd. 2015 European journal of endocrinology Vol.173 No.4

<P><B>Objective</B></P><P>Fine needle aspiration cytology (FNAC) and measurement of thyroglobulin (Tg) in needle washout (FNA-Tg) are recommended for the diagnosis of metastatic or recurrent lymph nodes (LNs) in differentiated thyroid cancer (DTC). However, the effect of serum Tg antibody (TgAb) on FNA-Tg levels still remains unclear in the preoperative setting. We analyze the interference of serum TgAb on FNA-Tg levels as proof of concept in the diagnostic advantage of serum TgAb combined with FNA-Tg.</P><P><B>Subjects and methods</B></P><P>A total of 370 suspicious cervical LNs from 273 patients with DTC were included. The primary tumor was confirmed as DTC on preoperative pathology in all patients. We performed FNA-Tg measurement and FNAC on suspicious LNs and evaluated the diagnostic performance of FNAC and FNA-Tg according to TgAb status. Final diagnoses were confirmed by histological examination of excised specimens or by follow-up ultrasonography for at least 6 months.</P><P><B>Results</B></P><P>Data from 273 subjects with suspicious 370 LNs were evaluated. Fifty-five LNs (14.9%) were from TgAb+ positive serum TgAb (TgAb+) patients. Serum Tg and FNA-Tg levels were significantly lower in patients with TgAb+ than in those with TgAb-negative (TgAb−). Final pathology confirmed 109 LNs (29.5%) asmalignant. Diagnostic performance of FNA-Tg at the same cutoff level was lower in the TgAb+ than TgAb− group. FNA-Tg cutoff levels determined by ROC curve were lower in the TgAb+ group.</P><P><B>Conclusion</B></P><P>The results suggested that the cutoff value of FNA-Tg should be lowered in suspicious LN before thyroidectomy in thyroid cancer patients with TgAb.</P>

Caffeine promotes premature chromosome condensation formation and in vitro development in porcine reconstructed embryos via a high level of maturation promoting factor activity during nuclear transfer.

Kawahara, Manabu,Wakai, Takuya,Yamanaka, Ken-Ichi,Kobayashi, Jin,Sugimura, Satoshi,Shimizu, Takashi,Matsumoto, Hiromichi,Kim, Jin-Hoi,Sasada, Hiroshi,Sato, Eimei BioScientifica 2005 Reproduction Vol.130 No.3

<P>When the nucleus in G0/G1 phase is transferred to an enucleated oocyte by nuclear transfer (NT), its nuclear envelope is broken, followed by condensation of chromosome structure by maturation promoting factor (MPF). This morphological remodeling of the transferred interphase nucleus seems to be essential for subsequent development of NT embryos. In this study, we treated porcine NT embryos with caffeine, which has been reported to increase MPF activity, to keep their MPF level high during NT. When 2.5 mM caffeine was added to the handling medium, the proportion of NT embryos showing condensed chromosome increased significantly (P < 0.05). In NT embryos treated with caffeine, the activity of p34(cdc2) kinase was significantly (P < 0.05) higher than in those without caffeine at 3 h post-injection. In addition, the rate of development to the blastocyst stage after activation was significantly (P < 0.05) higher in NT embryos treated with caffeine. These results indicate that caffeine treatment can increase not only the rate of chromosome condensation but also the developmental rate to the blastocyst stage of porcine NT embryos. This action is most likely due to the support/increase of MPF activity throughout the process of NT.</P>

Low PR in ER(+)/HER2(−) breast cancer: high rates of <i>TP53</i> mutation and high SUV

Ahn, Sung Gwe,Yoon, Chang Ik,Lee, Jae Hoon,Lee, Hye Sun,Park, So Eun,Cha, Yoon Jin,Cha, Chihwan,Bae, Soong June,Lee, Kyung-A,Jeong, Joon Bioscientifica Ltd 2018 Endocrine-related cancer Vol.26 No.2

<P>On the basis of <I>TP53</I> mutations and standardized uptake values (SUVs) from 18F-fluorodeoxyglucose positron emission tomography (18F-FDG-PET), we sought to enhance our knowledge of the biology underlying low progesterone receptor (PR) expression in estrogen receptor (ER)-positive/human epidermal growth factor receptor-2 (HER2)-negative tumors. This study included 272 patients surgically treated for ER-positive, HER2-negative breast cancer and who had undergone <I>TP53</I> gene sequencing. Of these, 229 patients also underwent 18F-FDG PET or PET/CT. Mutational analysis of exons 5–9 of the <I>TP53</I> gene was conducted using PCR amplification and direct sequencing. The SUVs were measured using 18F-FDG-PET scan images. Twenty-eight (10.3%) tumors had a somatic <I>TP53</I> mutation. The <I>TP53</I> mutation rate was significantly higher in low-PR tumors than in high-PR tumors (17.1% vs 7.9%, <I>P</I> = 0.039). Low-PR tumors had significantly higher median SUVs than high-PR tumors (<I>P</I> = 0.046). The multivariable analysis revealed that SUV and age remained independent variables associated with low PR expression. An adverse impact of low PR expression on recurrence-free survival was observed in the multivariable Cox regression hazard model. We provide clinical evidence that genetic alteration of the <I>TP53</I> gene and dysregulated glucose metabolism partly involve low PR expression in ER-positive and HER2-negative breast cancer.</P>

Evaluation of estrogenic potential by herbal formula, HPC 03 for <i>in vitro</i> and <i>in vivo</i>

Chang, Bo Yoon,Kim, Dae Sung,Kim, Hye Soo,Kim, Sung Yeon BioScientifica Ltd 2018 Reproduction Vol.155 No.2

<P>HPC 03 is herbal formula that consists of extracts from Angelica gigas, Cnidium officinale Makino and Cinnamomum cassia Presl. The present study evaluated the estrogenic potential of HPC 03 by using in vitro and in vivo models. The regulatory mechanisms of HPC 03 in estrogen-dependent MCF-7 cells were assessed. HPC 03 induced the proliferation of estrogen receptor-positive MCF-7 cells, and the proliferation was blocked by the addition of the estrogen antagonist tamoxifen. The estrogen receptora(alpha/beta) luciferase activities were significantly increased by HPC 03 treatment, which also increased the mRNA expression of the estrogen-responsive genes Psen2, Pgr and Ctsd. Also, we evaluated the ameliorative effects of HPC 03 on menopausal symptoms in ovariectomized rats. HPC 03 treatment in OVX rats significantly affected the uterine weight, increased the expression of estrogen-responsive genes Pgr and Psen2 in uterus, increased bone mineral density loss in the femur and inhibited body weight increase. Serum E2, collagen type 1 and osteocalcin were significantly increased, while serum LH, FSH and ALP were decreased compared with OVX rats. HPC 03 may be a promising candidate for the treatment of menopause, but further research is necessary to determine whether the observed effects also occur in humans.</P>

Identification and differential expression patterns of porcine <i>OCT4</i> variants

Hwang, Jae Yeon,Oh, Jong-Nam,Lee, Dong-Kyung,Choi, Kwang-Hwan,Park, Chi-Hun,Lee, Chang-Kyu BioScientifica 2015 Reproduction Vol.149 No.1

<P><I>OCT4</I> encoded by <I>POU5F1</I> has a crucial role of maintaining pluripotency in embryonic stem cells during early embryonic development and several <I>OCT4</I> variants have been identified in mouse and human studies. The objective of this study was to identify different variants of <I>OCT4</I> and analyze their expression patterns in preimplantation porcine embryos and various tissues. In this study, we showed that <I>POU5F1</I> transcribes its three variants, namely <I>OCT4A</I>, <I>OCT4B</I>, and <I>OCT4B1</I>. The <I>OCT4B</I> transcript consists of exons identical to the major form of the <I>OCT4</I> variant, <I>OCT4A</I>, with a differential N-terminal domain-coding exon. The structure of <I>OCT4B1</I> mRNA was the same as that of <I>OCT4B</I> mRNA, but harbored a cryptic exon. Based on these findings, the transcription levels were investigated and found that <I>OCT4B</I> and <I>OCT4B1</I> made up ∼20% among the variants in the embryonic stage and this indicates that <I>OCT4A</I> mRNA is dominantly expressed during preimplantation embryo development. In addition, <I>OCT4B</I> mRNA was detected in all tissues examined, while <I>OCT4A</I> and <I>OCT4B1</I> were detected only in testis but not in other tissues examined. <I>OCT4B1</I> showed inversely correlated expression with <I>SOX2</I> and <I>NANOG</I> expression. OCT4A protein was specifically localized to the nuclei, whereas OCT4B was mainly localized to the cytoplasm of the porcine embryos at the blastocyst stage. The findings of this study reveal that the porcine <I>OCT4</I> gene can potentially encode three variants (<I>OCT4A</I>, <I>OCT4B</I>, and <I>OCT4B1</I>), and they are differentially expressed and would have roles dissimilar between each other in preimplantation embryos and various adult tissues.</P>

Inhibition of DNA repair protein RAD51 affects porcine preimplantation embryo development

Jin, Zhe-Long,Shen, Xing-Hui,Shuang, Liang,Kwon, Jeong-woo,Seong, Min-Jeong,Kim, Nam-Hyung BioScientifica 2019 Reproduction Vol.157 No.3

<P>Homologous recombination (HR) plays a critical role in facilitating replication fork progression when the polymerase complex encounters a blocking DNA lesion, and it also serves as the primary mechanism for error-free DNA repair of double-stranded breaks. DNA repair protein RAD51 homolog 1 (RAD51) plays a central role in HR. However, the role of RAD51 during porcine early embryo development is unknown. In the present study, we examined whether RAD51 is involved in the regulation of early embryonic development of porcine parthenotes. We found that inhibition of RAD51 delayed cleavage and ceased development before the blastocyst stage. Disrupting RAD51 activity with RNAi or an inhibitor induces sustained DNA damage, as demonstrated by the formation of distinct γH2AX foci in nuclei of four-cell embryos. Inhibiting RAD51 triggers a DNA damage checkpoint by activating the ataxia telangiectasia mutated (ATM)-p53-p21 pathway. Furthermore, RAD51 inhibition caused apoptosis, reactive oxygen species accumulation, abnormal mitochondrial distribution and decreased pluripotent gene expression in blastocysts. Thus, our results indicate that RAD51 is required for proper porcine parthenogenetic activation (PA) embryo development.</P>

Spatial and temporal action of chicken primordial germ cells during initial migration

Soo Kang, Kyung,Chul Lee, Hyung,Jeong Kim, Hyun,Gun Lee, Hyo,Min Kim, Young,Jo Lee, Hong,Hyun Park, Young,Yeong Yang, Seo,Rengaraj, Deivendran,Sub Park, Tae,Yong Han, Jae BioScientifica 2015 Reproduction Vol.149 No.2

<P>In most animals, primordial germ cells (PGCs) originate from an extragonadal region and migrate across the embryo to the gonads, where they differentiate and function. During their migration, PGCs move passively by morphogenetic movement of the embryo or move actively through signaling molecules. To uncover the underlying mechanism of first-phase PGC migration toward the germinal crescent in chickens, we investigated the spatial and temporal action of PGCs during primitive streak formation. Exogenously transplanted PGCs migrated toward the anterior region of the embryo and the embryonic gonads when they were transplanted into the subgerminal cavity, but not into the posterior marginal zone, in Eyal–Giladi and Kochav stage X embryos. These results indicate that for passive migration toward the anterior region the initial location of PGCs should be the central region. Notably, although PGCs and DF-1 cells migrated passively toward the anterior region, only PGCs migrated to the germinal crescent, where endogenous PGCs mainly reside, by active movement. In a live-imaging experiment with green fluorescence protein-expressing transgenic embryos, exogenous PGCs demonstrated markedly faster migration when they reached the anterior one-third of the embryo, while somatic cells showed epiblast movement with constant speed. Also, migrating PGCs exhibited successive contraction and expansion indicating their active migration. Our results suggest that chicken PGCs use sequential passive and active forces to migrate toward the germinal crescent.</P>