Interaction of arylsulfatases A and B with maspin: A possible explanation for dysregulation of tumor cell metabolism and invasive potential of colorectal cancer

BACKGROUND Although it has been shown that arylsulfatases are lost in colorectal cancer(CRC)cell lines,their exact role in the carcinogenesis and behavior of this cancer was not elucidated.No data about the correlation between serum and immunohistochemical(IHC)level of arylsulfatases(ARSA,ARSB)in patients with CRC were published yet.AIM To evaluate the possible prognostic value of ARSA and/or ARSB in CRC,at circulating and protein levels.METHODS The present study included 45 consecutive patients who were prospectively diagnosed with CRC.For IHC stains(protein expression)ARSA,ARSB and maspin expression were quantified.For these markers,cytoplasmic expression was taken into account.For gene expression study,circulating mRNA was isolated from all patients,before surgery.A group of 45 healthy patients without inflammatory or tumor pathologies was used as control group.Reverse transcription and Taqman Gene Expression Array were used for ARSB gene expression.RESULTS The preoperative circulating RNA level of the ARSB gene was significantly decreased in patients with CRC(RQ<1),compared with the control group(RQ>1).A more significant decrease(RQ<0.5)occurred in ulcero-infiltrative maspinpositive adenocarcinomas,with a higher degree of tumor budding,diagnosed in locally advanced stages(pT3/4).ARSA/maspin immunopositivity indicated a higher risk for lymph node metastasis,while triple positivity for maspin/ARSA/ARSB and ARSB gene expression level<0.5 were indicators of CRC aggressive behavior,independent of lymph node status.CONCLUSION The significant independent negative prognostic factors of CRC are the ulceroinfiltrative aspect,high budding degree,triple positivity for maspin,ARSA and ARSB,and low ARSB gene expression.

Subcellular expression of maspin – from normal tissue to tumor cells

Maspin or SerpinB5, a member of the serine protease inhibitor family, was shown to function as a tumor suppressor, especially in carcinomas. It seems to inhibit invasion, tumor cells motility and angiogenesis, and promotes apoptosis. Maspin can also induce epigenetic changes such as cytosine methylation, de-acetylation,chromatin condensation, and histone modulation. In this review, a comprehensive synthesis of the literature was done to present maspin function from normal tissues to pathologic conditions. Data was sourced from MEDLINE and PubMed. Study eligibility criteria included: Published in English, between 1994 and 2019, specific to humans, and with full-text availability. Most of the 118 studies included in the present review focused on maspin immunostaining and m RNA levels. It was shown that maspin function is organ-related and depends on its subcellular localization. In malignant tumors, it might be downregulated or negative(e.g., carcinoma of prostate, stomach, and breast) or upregulated(e.g.,colorectal and pancreatic tumors). Its subcellular localization(nuclear vs cytoplasm), which can be proved using immunohistochemical methods, was shown to influence both tumor behavior and response to chemotherapy.Although the number of maspin-related papers increased, the exact role of this protein remains unknown, and its interpretation should be done with extremely high caution.

DNA extraction from paraffin embedded colorectal carcinoma samples: A comparison study of manual vs automated methods,using four commercially kits

BACKGROUND Nucleic acid isolation from formalin-fixed, paraffin-embedded tissue(FFPET)samples is a daily routine in molecular pathology laboratories, but extraction from FFPET is not always easily achieved. Choosing the right extraction technique is key for further examinations.AIM To compare the performance of four commercially available kits used for DNA extraction in routine practice.METHODS DNA isolation was performed on 46 randomly selected formalin-fixed, paraffinembedded(FFPE) colorectal adenocarcinoma(CRC) surgical specimens. Four commercially available extraction kits were used: two for manual DNA extraction(the Pure Link Genomic DNA Mini Kit from Invitrogen and the High Pure FFPE DNA Isolation Kit from Roche) and two for automated DNA extraction(the i Prep Genomic DNA Kit from Invitrogen and the Magna Pure LC DNA Isolation Kit from Roche). The DNA concentration and quality(odds ratio) among the four systems were compared. The results were correlated with the clinicopathological aspects of CRC cases: age, gender, localization, macro-and microscopic features,lymph node metastases, and the lymph node ratio.RESULTS The highest DNA concentration was obtained using the manual kits: 157.24 ±62.99 ng/μL for the Pure Link Genomic DNA Mini Kit and 86.64 ng/μL± 43.84 for the High Pure FFPE DNA Isolation Kit(P < 0.0001). Lower concentrations were obtained with automated systems: 20.39 ± 21.19 ng/μL for the Magna Pure LC DNA Isolation Kit and 8.722 ± 6.408 ng/μL for the i Prep Genomic DNA Kit,with differences between the systems used(P < 0.0001). The comparison between age, gender, tumor localization, pT or pN stage and the lymph node ratio indicated no statistically significant difference in DNA concentration using any of the nucleic acid isolation kits. DNA concentration was influenced by the macroscopic features and grade of differentiation. A higher DNA concentration was obtained for well-differentiated polypoid colorectal adenocarcinomas(CRCs), compared with undifferentiated ulcero-infiltrative carcinomas,irrespective of the kit used.CONCLUSION For research or diagnosis that needs high DNA concentrations, manual methods of DNA isolation should be used. A higher amount of DNA can be obtained from polypoid-type differentiated CRCs. Automated systems confer comfort and a lower amount of DNA that is, however, sufficient for classic polymerase chain reaction(PCR) and real-time quantitative PCR molecular examinations. All four commercially available kits can be successfully used in daily practice.

Cystic low-grade collecting duct renal carcinoma with liver compression—A challenging diagnosis and therapy: A case report

BACKGROUND A collecting duct carcinoma is a very rare, malignant renal epithelial tumor.Distant metastases are present in one third of cases at the time of diagnosis. It is known to have a poor prognosis.CASE SUMMARY A 42-year-old male was sent to our surgery clinic for removal of a 119.2 mm ×108.3 mm encapsulated cystic mass, which was localized in the 8th segment of the right liver lobe. The lesion was first identified on ultrasonography. A computed tomography scan confirmed the presence of a Bosniak type Ⅲ cystic lesion,which affected the liver and convexity of the right kidney. Surgical intervention involved a right nephrectomy, with removal of the cystic mass. The patient was mobilized on the first postoperative day and was discharged after 7 d. The histological and immunohistochemical examination revealed a low-grade collecting duct renal carcinoma, which is a rare variant of papillary carcinoma,with low malignant potential. The patient did not receive chemotherapy and after 21 mo of follow-up, a radiological examination and laboratory analyses showed normal aspects. No relapse or other complications were reported.CONCLUSION To manage renal tumors properly, a correct histopathological diagnosis is crucial,as is early diagnosis and correct surgical treatment.