Frequent alterations of SLIT2–ROBO1–CDC42 signalling pathway in breast cancer: clinicopathological correlation

The aim of the study was to understand the role of SLIT2–ROBO1/2–CDC42 signalling pathways in development of breast cancer (BC). Primary BC samples (n=150), comprising of almost equal proportion of four subtypes were tested for molecular alterations of SLIT2, ROBO1, ROBO2 and CDC42, the key regulator genes of this pathway. Deletion and methylation frequencies of the candidate genes were seen in the following order: deletion, SLIT2 (38.6%) >ROBO1 (30%) >ROBO2 (7.3%); methylation, SLIT2 (63.3%) >ROBO1 (26.6%) >ROBO2 (9.3%). Majority (80%, 120/150) of the tumours showed alterations (deletion/methylation) in at least one of the candidate genes. Overall, alterations of the candidate genes were as follows: SLIT2, 75.3% (101/150); ROBO1, 45.3% (68/150); ROBO2, 15.3% (23/150). Significantly, higher alteration of SLIT2 locus was observed in triple negative breast cancer (TNBC) over HER2 subtype (P=0.0014). Similar trend is also seen in overall alterations of SLIT2 and/or ROBO1, in TNBC than HER2 subtype (P=0.0012); of SLIT2 and/or ROBO2 in TNBC than luminal A (P=0.014) and HER2 subtype (P=0.048). Immunohistochemical analysis of SLIT2, ROBO1/2 showed reduced expression, concordant with their molecular alterations. Also, high expression of total CDC42 (49/52; 94.2%) and reduced expression of phospho Serine-71 CDC42 (41/52; 78.8%) was observed. Coalterations of SLIT2 and/or ROBO1, SLIT2 and/or ROBO2 had significant association with reduced expression of phospho Serine-71 CDC42 (P=0.0012–0.0038). Alterations of SLIT2 and/or ROBO1, reduced expression of phospho Serine-71 CDC42 predicted poor survival of BC patients. Results indicate the importance of SLIT2–ROBO1–CDC42 signalling pathway in predicting tumour progression.     

印度加尔各答市区外围一地的露尾甲物种组成、多度和季节性发生(英文)

【目的】印度大部分露尾甲在腐烂的水果和蔬菜上大量发生,其种群在一年中表现出明显的季节性波动。据推测,露尾甲种群很大程度上依赖于温度、湿度和降雨之类的环境因子。【方法】本研究调查了2013-2015年印度加尔各答市区外围一地Garia的露尾甲物种组成、季节性发生和种群结构,记录了其活跃时期、季节性多度和影响其发生的因素。【结果】调查期间在调查地共发现数目不等的6个物种。其中最常见露尾甲为Urophorus humeralis,它是个体数量最多的物种且在一年中几乎所有月份均有发生;其他常见物种为Epuraea ocularis和E. luteola。不同物种在食物发酵的连续阶段进入诱捕器中。最初12 h被捕获的是Epuraea 属的种类,而在诱捕器中食物严重腐烂的后续阶段发现最多的是U. humeralis。在合适范围的气温(22~29℃)和相对湿度(82.5%~86%)下,物种丰富度最高,表明这些环境变量对露尾甲种群具有重要影响。【结论】加尔各答主要水果和蔬菜在季风后季节种植,在季风后季节取食这些作物的露尾甲发生量(物种丰富度和多度)最高。这一研究结果可能有助于制定针对这些甲虫的有效田间治理策略。     

Selective and Synergistic Activity of L-S,R-Buthionine Sulfoximine on Malignant Melanoma Is Accompanied by Decreased Expression of Glutathione-S-Transferase

L-buthionine-S,R-sulfoximine (BSO) selectively inhibits glutathione (GSH) synthesis. Malignant melanoma may be uniquely dependent on GSH and its linked enzymes, glutathione S-transferase (GST) and GSH-peroxidase, for metabolism of reactive orthoquinones and peroxides produced during melanin synthesis. We compared the in vitro effects of BSO on melanoma cell lines and fresh melanoma specimens (n = 118) with breast and ovarian cell lines and solid tumors (n = 244). IC₅₀ values (μM) for BSO on melanoma, breast and ovarian tumor specimens were 1.9, 8.6, and 29, respectively. The IC₉₀ for melanoma was 25.5 μM, a level 20-fold lower than steady state levels achieved clinically. The sensitivity of individual specimens of melanoma correlated with their melanin content (r = 0.63). BSO synergistically enhanced BCNU activity against melanoma cell lines and human tumors. We followed GSH levels, GST enzyme activity, GST isoenzyme profiles and mRNA levels after BSO. BSO (50 μM) treatment for 48 hr resulted in a 95% decrease in ZAZ and M14 melanoma cell line GSH levels, and a 60% decrease in GST enzyme activity. GST-μ. protein and mRNA levels were significantly reduced in both cell lines. GST expression was unaffected. These data suggest that BSO action on melanoma may be related to GSH depletion, diminishing the capacity to scavenge toxic metabolites produced during melanin synthesis. We report here for the first time that BSO enhancement of alkylator action may be related in part to down regulation of GST. BSO may be a clinically useful adjunct in the treatment of malignant melanoma.