Overexpression of N-Myc Downstream-Regulated Gene 2 (NDRG2) Regulates the Proliferation and Invasion of Bladder Cancer Cells In Vitro and In Vivo

N-Myc downstream-regulated gene 2 (NDRG2) is a candidate tumor suppressor gene, which plays an important role in controlling tumor growth. The aim of this study was to investigate the expression of NDRG2 gene in bladder cancer (BC) tissues and several bladder cancer cell lines, and to seek its clinical and pathological significance. Ninety-seven bladder carcinoma and 15 normal bladder tissue sections were analyzed retrospectively with immunohistochemistry. The human bladder cancer cell line T24 was infected with LEN-NDRG2 or LEN-LacZ. The effects of NDRG2 overexpression on T24 cells and T24 nude mouse xenografts were measured via cell growth curves, tumor growth curves, flow cytometric analysis, western blot and Transwell assay. NDRG2 was highly expressed in normal bladder tissue, but absent or rarely expressed in cacinomatous tissues (χ2=8.761, p < 0.01). The NDRG2 level was negatively correlated with tumor grade and pathologic stage(r=-0.248, p < 0.05), as well as increased c-myc level (r=-0.454, p< 0.001). The expression of NDRG2 was low in the three BC cell lines. T24 cells infected with LEN-NDRG2 showed inhibition of proliferation both in vitro and in vivo, and NDRG2 overexpression can inhibit tumor growth and invasion in vitro

Factors Associated with Gender-Affirming Surgery and Age of Hormone Therapy Initiation Among Transgender Adults

Abstract Purpose: Gender-affirming surgeries and hormone therapy are medically necessary treatments to alleviate gender dysphoria; however, significant gaps exist in the research and clinical literature on surgery utilization and age of hormone therapy initiation among transgender adults. Methods: We conducted a retrospective review of electronic health record data from a random sample of 201 transgender patients of ages 18–64 years who presented for primary care between July 1, 2010 and June 30, 2015 (inclusive) at an urban community health center in Boston, MA. Fifty percent in our analyses were trans masculine (TM), 50% trans feminine, and 24% reported a genderqueer/nonbinary gender identity. Regression models were fit to assess demographic, gender identity-related, sexual history, and mental health correlates of gender-affirming surgery and of age of hormone therapy initiation. Results: Overall, 95% of patients were prescribed hormones by their primary care provider, and the mean age of initiation of masculinizing or feminizing hormone prescriptions was 31.8 years (SD=11.1). Younger age of initiation of hormone prescriptions was associated with being TM, being a student, identifying as straight/heterosexual, having casual sexual partners, and not having past alcohol use disorder. Approximately one-third (32%) had a documented history of gender-affirming surgery. Factors associated with increased odds of surgery were older age, higher income levels, not identifying as bisexual, and not having a current psychotherapist. Conclusion: This study extends our understanding of prevalence and factors associated with gender-affirming treatments among transgender adults seeking primary care. Findings can inform future interventions to expand delivery of clinical care for transgender patients

Mining the Plant-Herbivore Interface with a Leafmining Drosophila of Arabidopsis

Experimental infections of Arabidopsis thaliana (Arabidopsis) with genomically characterized plant pathogens such as Pseudomonas syringae have facilitated the dissection of canonical eukaryotic defence pathways and parasite virulence factors. Plants are also attacked by herbivorous insects, and the development of an ecologically relevant genetic model herbivore that feeds on Arabidopsis will enable the parallel dissection of host defence and reciprocal resistance pathways such as those involved in xenobiotic metabolism. An ideal candidate is Scaptomyza flava, a drosophilid fly whose leafmining larvae are true herbivores that can be found in nature feeding on Arabidopsis and other crucifers. Here, we describe the life cycle of S. flava on Arabidopsis and use multiple approaches to characterize the response of Arabidopsis to S. flava attack. Oviposition choice tests and growth performance assays on different Arabidopsis ecotypes, defence-related mutants, and hormone and chitin-treated plants revealed significant differences in host preference and variation in larval performance across Arabidopsis accessions. The jasmonate and glucosinolate pathways in Arabidopsis are important in mediating quantitative resistance against S. flava, and priming with jasmonate or chitin resulted in increased resistance. Expression of xenobiotic detoxification genes was reduced in S. flava larvae reared on Arabidopsis jasmonate signalling mutants and increased in plants pretreated with chitin. These results and future research directions are discussed in the context of developing a genetic model system to analyse insect–plant interactions.Organismic and Evolutionary Biolog

<i>NDRG2</i> overexpression inhibits proliferation of human bladder cancer cells.

<p>(A and B) Colony formation assay. Upregulation of <i>NDRG2</i> inhibits colony formation. (C) The cell growth curves of T24 cells by MTT method. All the assays were repeated independently for at least three times. The results are shown as the mean ± SD (*=p<0.001).</p

The influences of <i>NDRG2</i> overexpression on T24 cells infected with lentivirus.

<p>(A) The effect of <i>NDRG2</i> overexpression on G1 cell cycle and apoptosis regulators as assayed by western blot. (B) Relative quantification of protein expression, normalized to GAPDH levels. The figure shows the tendency of each group as indicated. All the assays were repeated independently for at least three times. The results are shown as the mean ± SD (** p<0.01)..</p

The expression of <i>NDRG2</i> and c-Myc protein in bladder carcinoma tissues detected with immunohistochemistry staining

<p>(A) The expression levels of <i>NDRG2</i> protein decrease while the degree of malignancy of bladder carcinoma increases (x400)(1). Normal bladder tissue; (2) Bladder papilloma(T0-Ta); (3) High-level bladder cancer(T2-T4). (B) The expression levels of c-Myc protein increases while the degree of malignancy of bladder carcinoma increases (x400)(4). Normal bladder tissue; (5) Bladder papilloma(T0-Ta); (6) High-level bladder cancer(T2-T4). The sections of (B) (4–6) originated from the same paraffin blocks as (A) (1–3) respectively.</p

FCA of cell cycle arrest and apoptosis induced by overexpression of <i>NDRG2</i>.

<p>(A) T24cells infected with LEN-<i>NDRG2</i> lentivirus were more obviously arrested in G0/G1cycle. (B) Percentages of apoptotic cells in LEN-<i>NDRG2</i> groups were greater than in the other two groups(1–3). 48 h after the infection (4–6); 72 h after the infection; (1 and 4) blank control (PBS); (2 and 5) LEN-LacZ; (3 and 6) LEN-<i>NDRG2</i>. All the assays were repeated independently for at least three times. The results are shown as the mean ± SD.</p

Differential expression of <i>NDRG2</i> among bladder cancer cell lines

<p>(A) Real-time PCR analysis of <i>NDRG2</i> mRNA expression. (B) Expression level of <i>NDRG2</i> protein as assayed by western blot. All the assays were repeated independently for at least three times. The results are shown as the mean ± SD (** p<0.001)..</p

The influences of <i>NDRG2</i> overexpression on T24 cells infected with lentivirus <i>in vivo</i>.

<p>(A) The effect of <i>NDRG2</i> overexpression on cell cycle, apoptosis and metastasis regulators as assayed by western blot. (B) Relative quantification of protein expression, normalized to GAPDH levels. Western blots were analyzed with Kodak Digital Science one-dimensional software. The figure shows the tendency of each group as indicated. All the assays were repeated independently for at least three times. The results are shown as the mean ± SD (* p<0.01).</p