GPR125 (ADGRA3) is an autocleavable adhesion GPCR that traffics with Dlg1 to the basolateral membrane and regulates epithelial apicobasal polarity.

The adhesion family of G protein-coupled receptors (GPCRs) is defined by an N-terminal large extracellular region that contains various adhesion-related domains and a highly-conserved GPCR-autoproteolysis-inducing (GAIN) domain, the latter of which is located immediately before a canonical seven-transmembrane domain. These receptors are expressed widely and involved in various functions including development, angiogenesis, synapse formation, and tumorigenesis. GPR125 (ADGRA3), an orphan adhesion GPCR, has been shown to modulate planar cell polarity in gastrulating zebrafish, but its biochemical properties and role in mammalian cells have remained largely unknown. Here, we show that human GPR125 likely undergoes cis-autoproteolysis when expressed in canine kidney epithelial MDCK cells and human embryonic kidney HEK293 cells. The cleavage appears to occur at an atypical GPCR proteolysis site within the GAIN domain during an early stage of receptor biosynthesis. The products, i.e., the N-terminal and C-terminal fragments, seem to remain associated after self-proteolysis, as observed in other adhesion GPCRs. Furthermore, in polarized MDCK cells, GPR125 is exclusively recruited to the basolateral domain of the plasma membrane. The recruitment likely requires the C-terminal PDZ-domain-binding motif of GPR125 and its interaction with the cell polarity protein Dlg1. Knockdown of GPR125 as well as that of Dlg1 results in formation of aberrant cysts with multiple lumens in Matrigel 3D culture of MDCK cells. Consistent with the multilumen phenotype, mitotic spindles are incorrectly oriented during cystogenesis in GPR125-KO MDCK cells. Thus, the basolateral protein GPR125, an autocleavable adhesion GPCR, appears to play a crucial role in apicobasal polarization in epithelial cells.

Novel mobilization of the medial approach without changing the position for robotic right hemicolectomy.

Conventional laparoscopic or robotic surgery for right-sided colon cancer often requires intraoperative repositioning and removal of the bowel. Changing positions during robotic surgery can be troublesome and robotic removal of the small intestine carries a risk of unexpected injury because robotic devices have a strong grasping force and no sense of touch. Herein, we introduce a novel mobilization of the medial approach without changing the position for robotic right hemicolectomy. Using this technique, mobilization is performed in counterclockwise succession, allowing all mobilizations and bowel removal to be completed sequentially, without positional change.

Long noncoding RNA NORAD regulates transforming growth factor-ß signaling and epithelial-to-mesenchymal transition-like phenotype.

Long noncoding RNAs are involved in a variety of cellular functions. In particular, an increasing number of studies have revealed the functions of long noncoding RNA in various cancers; however, their precise roles and mechanisms of action remain to be elucidated. NORAD, a cytoplasmic long noncoding RNA, is upregulated by irradiation and functions as a potential oncogenic factor by binding and inhibiting Pumilio proteins (PUM1/PUM2). Here, we show that NORAD upregulates transforming growth factor-ß (TGF-ß) signaling and regulates TGF-ß-induced epithelial-to-mesenchymal transition (EMT)-like phenotype, which is a critical step in the progression of lung adenocarcinoma, A549 cells. However, PUM1 does not appear to be involved in this process. We thus focused on importin ß1 as a binding partner of NORAD and found that knockdown of NORAD partially inhibits the physical interaction of importin ß1 with Smad3, inhibiting the nuclear accumulation of Smad complexes in response to TGF-ß. Our findings may provide a new mechanism underlying the function of NORAD in cancer cells.

Short-term outcomes of intracorporeal versus extracorporeal anastomosis in laparoscopic surgery for right-sided colon cancer: A propensity score-matched study.

INTRODUCTION: Recently, intracorporeal anastomosis (IA) has been attracting attention. We aimed to compare the short-term outcomes of IA and extracorporeal anastomosis (EA) in laparoscopic surgery for right-sided colon cancer, after propensity score matching. METHODS: We retrospectively reviewed 404 consecutive patients with right-sided primary colon cancer between January 2019 and July 2021, 359 of whom underwent laparoscopic surgery. We classified them into IA (n = 72) and EA (n = 287) groups. Propensity score matching analysis was performed, and the matched groups were compared. RESULTS: The IA group had a longer operation time and shorter time to first flatus, passage of stool, and oral intake. There were no differences in blood loss, postoperative complications, and postoperative hospital stay between the groups. The IA group had a higher inflammatory response in the laboratory data on postoperative day 1 compared to the EA group; however, there were no differences in the incidence of abdominal or surgical site infection (SSI). The IA group had a longer distal resection margin, and there were no peritoneal recurrences in either group. CONCLUSION: In the IA group, patients had earlier bowel recovery and a longer distal resection margin; however, other postoperative clinical outcomes were comparable. Although there was a higher postoperative inflammatory response in IA, there was no significant difference in postoperative complications, including SSI and intra-abdominal infection. Although long-term outcomes are not yet available, IA could be a useful procedure.

Laparoscopic resection for a relapsed presacral epidermoid cyst penetrating the ischiorectal fossa.

An epidermoid cyst occurring in the presacral region is a rare congenital cystic tumor. Complete surgical excision is recommended because of the possibility of infection and malignancy. Depending on the size and location, several approaches, such as anterior, posterior, combined, and laparoscopic approaches, have been described. Here, we have reported a rare case of relapsed epidermoid cyst after emergency laparotomy and transvaginal drainage. Although the cyst penetrated the levator ani muscle and was densely adhered to the vagina and rectum, complete laparoscopic resection was successfully performed. This approach is a useful technique as it is minimally invasive and minimizes tissue destruction of the pelvic floor.

ZEB1-regulated inflammatory phenotype in breast cancer cells.

Zinc finger E-box binding protein 1 (ZEB1) and ZEB2 induce epithelial-mesenchymal transition (EMT) and enhance cancer progression. However, the global view of transcriptional regulation by ZEB1 and ZEB2 is yet to be elucidated. Here, we identified a ZEB1-regulated inflammatory phenotype in breast cancer cells using chromatin immunoprecipitation sequencing and RNA sequencing, followed by gene set enrichment analysis (GSEA) of ZEB1-bound genes. Knockdown of ZEB1 and/or ZEB2 resulted in the downregulation of genes encoding inflammatory cytokines related to poor prognosis in patients with cancer, including IL6 and IL8, therefore suggesting that ZEB1 and ZEB2 have similar functions in terms of the regulation of production of inflammatory cytokines. Antibody array and ELISA experiments confirmed that ZEB1 controlled the production of the IL-6 and IL-8 proteins. The secretory proteins regulated by ZEB1 enhanced breast cancer cell proliferation and tumor growth. ZEB1 expression in breast cancer cells also affected the growth of fibroblasts in cell culture, and the accumulation of myeloid-derived suppressor cells in tumors in vivo. These findings provide insight into the role of ZEB1 in the progression of cancer, mediated by inflammatory cytokines, along with the initiation of EMT.