Lysophosphatidic acid receptor‐3 increases tumorigenicity and aggressiveness of rat hepatoma RH7777 cells

Lysophosphatidic acid (LPA), which interacts with G protein‐coupled transmembrane LPA receptors exhibits several biological effects, such as cell proliferation, migration, and differentiation. Recently, it has been reported that alteration of LPA receptor genes occurs in several cancer cells. In this study, to assess the biological role of LPA receptor‐3 (LPA₃) in the pathogenesis of tumor cells, we generated the Lpar3‐expressing cells (RHa3B12 and RHa3G8) from rat hepatoma RH7777 cells, and examined their abilities of cell migration and tumorigenicity, compared with the Lpar3‐unexpressing cells. In cell motility and invasion assays, RHa3B12 and RHa3G8 cells showed significantly higher intrinsic activity without LPA treatment than control RH7777AB cells. LPA treatment further increased cell motility and invasion of these cells. The cell motility of RHa3B12 and RHa3G8 cells stimulated by LPA treatment was significantly suppressed by pretreatment with inhibitors of Gi or Gq proteins. In a soft agar assay, the large sized colonies were formed in RHa3B12 and RHa3G8 cells, but not in RH7777AB cells. The cell survival of RHa3G8 cells treated with cisplatin (CDDP) or doxorubicin (DOX) was higher than that of RH7777AB cells, correlating with the elevated expression levels of multidrug‐resistance related genes, Mdr1a, Mdr1b, and Gstp1. These results suggest that LPA₃ may be involved in progression and aggressiveness of rat hepatoma RH7777 cells. © 2011 Wiley Periodicals, Inc.     

Simple, sensitive and reliable in vivo assays to evaluate the estrogenic activity of endocrine disruptors

Purpose

We compared three in vivo assays, determining changes of body weight, and uterotropic and vaginal cytology assays, for the evaluation of estrogenic activity of an estrogen disrupting compound, Pueraria mirifica (PM), in comparison with 17β-estradiol (E).

Methods

Female rats were ovariectomized and gavaged with distilled water, 0.01, 0.1, 1, 10 and 20 mg/kg BW/day of E and 100 and 1,000 mg/kg BW/day of PM for 14 days. Body weights were measured weekly, and vaginal epithelium cells were monitored daily. The uterus was dissected at the end of the treatment period, weighed and examined for histomorphometry.

Results

There were a decrease in body weight and an increase in uterine weight, uterine, endometrium and myometrium areas, uterine gland numbers, and percent of cornified cell which were dependent on doses of E and PM treatments.

Conclusions

Of the three assays proposed, although all are reliable and had critical read-out, measurements of body and uterine weights is likely convenient and simple, but the uterotropic assay needs to kill the animals. Vaginal cytology assay appears most promising for sensitivity and shortening the duration of the assay. Compared to those of E, the estrogenic activity of PM at concentrations of 100 and 1,000 mg/kg BW was in the range of 14 to >20 mg/kg BW.     

Neurodevelopmental and Behavioral Outcome of Very Low Birth Weight Babies at Corrected Age of 2 Years

Objective  

Neurodevelopmental and behavioral assessment of very low birth weight babies (VLBW) at corrected age (CA) of 2 years.     

Study of the poliovirus receptor related-1 gene in Thai patients with non-syndromic cleft lip with or without cleft palate

Non-syndromic cleft lip with or without cleft palate (CL/P) has a complex etiology with several genetic and environmental factors playing a role. The poliovirus receptor related-1 gene (PVRL1) has been shown to underlie a syndromic form of CL/P and, in some populations, contribute to non-syndromic CL/P. To investigate whether mutations in PVRL1 play a part in the formation of non-syndromic CL/P in the Thai population, 100CL/P patients were analyzed for mutations in PVRL1 by polymerase chain reaction amplification and direct sequencing of all the coding regions of its alpha isoform. Of this series of patients, one was found to be heterozygous for 1183G>A in exon 6, expected to result in the substitution of a valine by a methionine at position 395 (V395M). This mutation was not found in 200 unaffected Thai control individuals. The valine position is conserved across all known mammalian PVRL1 sequences. In conclusion, a novel non-synonymous PVRL1 mutation was found in a Thai patient with non-syndromic CL/P, suggesting a possible etiologic role of PVRL1 in non-syndromic CL/P across different populations.     

Oxidized high mobility group B‐1 enhances metastability of colorectal cancer via modification of mesenchymal stem/stromal cells

High mobility group box‐1 (HMGB1) is known to be a chemotactic factor for mesenchymal stem/stromal cells (MSCs), but the effect of post‐translational modification on its function is not clear. In this study, we hypothesized that differences in the oxidation state of HMGB1 would lead to differences in the function of MSCs in cancer. In human colorectal cancer, MSCs infiltrating into the stroma were correlated with liver metastasis and serum HMGB1. In animal models, oxidized HMGB1 mobilized three‐fold fewer MSCs to subcutaneous tumors compared with reduced HMGB1. Reduced HMGB1 inhibited the proliferation of mouse bone marrow MSCs (BM‐MSCs) and induced differentiation into osteoblasts and vascular pericytes, whereas oxidized HMGB1 promoted proliferation and increased stemness, and no differentiation was observed. When BM‐MSCs pretreated with oxidized HMGB1 were co‐cultured with syngeneic cancer cells, cell proliferation and stemness of cancer cells were increased, and tumorigenesis and drug resistance were promoted. In contrast, co‐culture with reduced HMGB1‐pretreated BM‐MSCs did not enhance stemness. In an animal orthotopic transplantation colorectal cancer model, oxidized HMGB1, but not reduced HMGB1, promoted liver metastasis with intratumoral MSC chemotaxis. Therefore, oxidized HMGB1 reprograms MSCs and promotes cancer malignancy. The oxidized HMGB1–MSC axis may be an important target for cancer therapy.     

Outcome of Reoperation for Local Recurrence Following En Bloc Resection for Bone Giant Cell Tumor of the Extremity

En bloc resection is typically performed to treat giant cell tumors of bone (GCTB), particularly when curettage can be challenging owing to extensive bone cortex destruction with soft tissue extension. Few reports have addressed the clinical outcomes after reoperation for local recurrence in patients with GCTB who underwent en bloc resection. In this multicenter retrospective study, we investigated local recurrence, distant metastasis, malignant transformation, mortality, and limb function in patients treated for local recurrence following en bloc resection for GCTB. Among 205 patients who underwent en bloc resection for GCTB of the extremities between 1980 and 2021, we included 29 with local recurrence. En bloc resection was performed for large tumors with soft tissue extension, pathological fractures with joint invasion, complex fractures, and dispensable bones, such as the proximal fibula and distal ulna. Local re-recurrence, distant metastasis, malignant transformation, and mortality rates were 41.4% (12/29), 34.5% (10/29), 6.9% (2/29), and 6.9% (2/29), respectively. The median Musculoskeletal Tumor Society score was 26 (interquartile range, 23–28). The median follow-up period after surgery for local recurrence was 70.1 months (interquartile range, 40.5–123.8 months). Local recurrence following en bloc resection for GCTB could indicate an aggressive GCTB, necessitating careful follow-up.     

Studies on azabicyclo systems: syntheses and spasmolytic activity of analogues of 9-methyl-3,9-diazabicyclo[4.2.1]nonane and 10-methyl-3,10-diazabicyclo[4.3.1]decane

The spasmolytic activity of the lactams, 9-methyl-3,9-diazabicyclo[4.2.1]nonan-4-one 1 and 10-methyl-3,10-diazabicyclo[4.3.1]decan-4-one 2 and their reduction products, 9-methyl-3,9-diazabicyclo[4.2.1]nonane 3 and 10-methyl-3,10-diazabicyclo[4.3.1]decane 4 are described. Syntheses and spasmolytic effects of new amides and sulfonamides of the amines 3 and 4 are also reported. 3 possessed specific anti-serotonin activity. Amides of 3 with aromatic acids resulted in specific anti-histaminic compounds, whereas amides of 4 with aliphatic acids showed spasmogenic activity. A number of amides and sulfonamides showed non-specific spasmolytic activity.     

Reconstruction after Talar Tumor Resection: A Systematic Review

This systematic review investigated the functional outcomes and complications of reconstruction methods after talar tumor resection. A systematic search of PubMed, Embase, and the Cochrane Central Register of Controlled Trials databases identified 156 studies, of which 20 (23 patients) were ultimately included. The mean Musculoskeletal Tumor Society scores in the groups reconstructed using tibiocalcaneal fusion (n = 17), frozen autograft (n = 1), and talar prosthesis (n = 5) were 77.6 (range 66–90), 70, and 90 (range 87–93), respectively. Regarding complications, sensory deficits were observed in one patient (6%) and venous thrombosis in two patients (12%) in the tibiocalcaneal fusion group, while osteoarthritis was observed in one patient (100%) in the frozen autograft group. No complications were observed in the talar prosthesis group. Reconstruction with talar prosthesis seems preferable to conventional tibiocalcaneal fusion after talar tumor resection because it offers better function and fewer complications. However, as this systematic review included only retrospective studies with a small number of patients, its results require re-evaluation in future randomized controlled trials with larger numbers of patients.