Effects of phillyrin on VEGF and endostatin expression in Lewis lung carcinoma

AIM: To investigate the effects of phillyrin on vascular endothelial growth factor (VEGF) and endostatin expression in lung tumor tissues isolated from Lewis lung carcinoma. METHODS: The expression of VEGF and endostatin in control individuals and the patients with lung cancer was determined by immunohistochemistry. In the animal experiment, 5 groups of animals were examined: control, tumor model, and tumor model with 3 different concentrations of phillyrin treatments. For preparation of transplanted tumor model, Lewis cells were subcutaneously injected into the right limb armpit of the nude mice. After that, phillyrin was administered via oral gavage once daily for 20 d at dose of 5 or 10 g/kg, or twice daily at 10 g/kg. Lung tumor tissues isolated from each group were observed by hematoxylin-eosin staining. VEGF and endostatin expression were examined by immunohistochemistry. RESULTS: VEGF expression was increased in lung tumor tissues as compared with normal and pericarcinous tissues, while endostatin expression was decreased. Phillyrin significantly inhibited the tumor size and tumor tissue density dose-dependently, which was accompanied with a decrease in VEGF expression and an increase in endostatin expression. CONCLUSION: Phillyrin inhibits the development of lung tumor through reducing VEGF expression and increasing endostatin expression.     

Preliminary exploration for effect of EphA2 on drug resistance of colorectal carcinoma cells

AIM: To investigate the effect of Eph receptor A2 (EphA2) on drug resistance of colorectal carcinoma cells and its possible mechanisms. METHODS: Real-time PCR and Western blot were used to detect the expression of EphA2 at mRNA and protein levels in LoVo and LoVo/5-FU cells. EphA2 siRNA was transfected to down-regulate the EphA2 expression in LoVo/5-FU cells, and the drug sensitivity was calculated by CCK-8 assay. Meanwhile, cell migration and invasion were measured by wound healing assay and Transwell assay, and the protein levels of E-cadherin, β-catenin, N-cadherin, vimentin, Notch and Snail were determined by Western blot. RESULTS: The expression of EphA2 at both mRNA and protein levels was significantly up-regulated in LoVo/5-FU cells (P<0.05). Knockdown of EphA2 suppressed the cell viability, and migration and invasion abilities, but promoted drug sensitivity of LoVo/5-FU cells. Up-regulation of E-cadherin and β-catenin, and down-regulation of N-cadherin and vimentin were observed, indicating that the epithelial-mesenchymal transition (EMT) process was suppressed. Knockdown of EphA2 decreased the expression levels of Notch and Snail. CONCLUSION: Down-regulation of EphA2 partly reverses drug resistance of LoVo/5-FU cells. The mechanism may be related to suppressing cell growth, migration, invasion and EMT process via Notch/Snail signaling pathway.     

Feline primary nonhematopoietic malignant liver tumours: A multicenter retrospective study (2000–2021)

There is scant literature on primary nonhematopoietic malignant liver tumours (PMLT) in cats. In this retrospective study, medical data of 40 cats diagnosed with PMLT were reviewed over a period of 22 years (2000–2021). The most frequent epithelial tumours were hepatocellular (42.5%) and bile duct carcinomas (32.5%), only six (15%) cats had mesenchymal tumours. The median age was 13 years and clinical signs commonly included ano-/hyporexia (62.5%), apathy/lethargy (52.5%), weight loss (42.5%) and vomiting (35%). At initial diagnosis, metastases were confirmed in 1 (2.5%) and suspected in three (7.5%) cats. Massive was the most frequent morphology (75%). Most intrahepatic tumours were left-sided (54.2%) with the left medial lobe being primarily affected (25%). Extrahepatic tumours were rare (5%). In 34 (85%) cats, liver lobectomy was performed (surgery group), four (10%) were treated palliatively (non-surgery group), and two (5%) received no treatment. Intraoperative complications occurred in 11.8% with four (15.4%) postoperative deaths. Recurrence was detected in 28.6% at a median of 151 days (range, 79–684 days), while postoperative metastases were suspected in 21.4% at a median of 186 days (range, 79–479 days). The median survival time (MST) was significantly longer in cats of the surgery group (375 days) than in the non-surgery group (16 days) (p = .002). MST was 868 days for hepatocellular compared to 270 days for bile duct carcinomas (p = .06). In summary, liver lobectomy is associated with prolonged survival times and good prognosis in cats with hepatocellular, and an acceptable prognosis in cats with bile duct carcinoma.     

Description and efficacy of a response-based “QUAD” cyclical hypofractionated palliative-intent radiation protocol in dogs with macroscopic solid tumours: 108 cases

Palliative-intent radiation therapy can alleviate pain and clinical signs in dogs with cancer, but optimal fractionation scheme is unknown. The objective of this retrospective case series is to evaluate clinical benefit, objective response, adverse effects, and outcomes in 108 dogs with macroscopic solid tumours treated with a cyclical “QUAD” hypofractionated palliative-intent radiation therapy protocol. Median QUAD dose was 14 Gy (14–16 Gy). Median total dose was 28 Gy (14–48 Gy). Clinical benefit rate was 93%, with median onset of subjective palliation 21 days after the first QUAD, lasting a median of 134 days. Tumour volumetric objective response was assessed with CT prior to the third QUAD in 36 dogs, with stable disease in 24 dogs (67%) and partial response in 9 dogs (25%). Sinonasal and oral were the most common tumour locations in 32 and 30 dogs, respectively. Median progression-free survival was 153 days (95% CI 114–200). Median overall survival was 212 days (95% CI 152–259). Number of QUAD cycles completed, clinical benefit achieved, anti-inflammatory received, total radiation dose, time to maximum clinical benefit, and response duration were positively associated with progression-free and overall survival. Acute toxicities were observed in 15 dogs (14%) with 3 high-grade (grade 3) toxicities (3%). Low-grade (grade 1 and 2) late skin and ocular toxicities were observed in 31 dogs (29%), predominantly leukotrichia, alopecia, keratoconjunctivitis sicca, and cataracts. This report demonstrates that QUAD radiation is an alternative protocol to be considered for palliation of dogs with inoperable or advanced stage solid tumours.     

Prevalence and anatomopathological characterization of cutaneous squamous cell carcinomas with regional and distant metastases in dogs and cats: 20 cases (1985–2020)

The aim of this study was to assess the prevalence of regional and distant metastases from cutaneous squamous cell carcinomas (SCCs) in dogs (n = 11) and cats (n = 9) in a retrospective case series over 36 years (1985–2020), as well as to characterize its macroscopic aspects (location and size), degree of differentiation (well, moderately and poorly differentiated [WD, MD and PD, respectively]) and the rate of cell proliferation, by counting the AgNORs. Immunohistochemistry (IHC) was used to identify patterns of tumour migration and invasion (islands, ribbons, cords, small aggregates, individual cells [fusiform and amoeboid]) and to evaluate the intensity of desmoplasia and the amount of myofibroblasts. The prevalence of metastatic SCCs was 4.39% (21/478), being 3.8% in dog (12/309) and 5.3% in cat (9/169). Metastases affected lymph nodes in all dogs and 66% (6/9) of cats, and less frequently distant organs. Primary tumours predominantly affected the abdominal skin in dogs and the nasal planum in cats. Among the 20 cases, 52% were MDs, 34% were WDs, and 14% were PDs. Histological lesions suggestive of exposure to chronic solar radiation were present in 57% (8/14). The main patterns of tumour migration and invasion were islands for WD SCCs and individual cells for PD SCCs. MD SCCs had a mix of patterns. In cats, individual spindle cells were restricted to PDs. A marked desmoplastic reaction was more associated with PD SCCs and often with MDs. This study highlights that the prevalence of SCC metastases in dogs and cats is predominantly regional. The IHC was essential in the identification of individual fusiform keratinocytes, whose presence in surgical margins may represent a greater risk of recurrence. Although the presence of myofibroblasts was observed in all infiltrative and metastatic tumours, further studies evaluating these cells may be important to better understand their role in the tumour microenvironment of cutaneous SCCs with metastasis in dogs and cats.     

Effect of microtubule-destabilizing protein stathmin gene silencing on proliferation and apoptosis of nasopharyngeal carcinoma cell line 5-8F

AIM: To investigate the effects of stathmin gene silencing on nasopharyngeal carcinoma cell line 5-8F. METHODS: Double-strand siRNA targeting to stathmin gene was obtained by chemical synthesis and annealing, and was sub-cloned into the vector pGenesil-1.1. The plasmid was introduced into 5-8F cells by liposome-mediated transfection. The gene expression of stathmin, and the proliferation, morphology and apoptosis of the cells were analyzed by Western blotting, MTT assay and flow cytometry. RESULTS: The cell suppression rate in stathmin gene silencing group was (53.01?1.12)%, significantly higher than that in transfection reagent group and in negative control group. The cell apoptotic rate in stathmin gene silencing group was (8.75?0.67)%, also significantly higher than that in transfection reagent group and in negative control group (P<0.05). CONCLUSION: Silencing of stathmin gene in nasopharyngeal carcinoma cells inhibits the cell proliferation and induces cell apoptosis.     

Tetrandrine enhances radiosensitivity of human nasophayrngeal carcinoma cell lines

AIM: To study whether tetrandrine (Tet) enhances the radiosensitivity of human nasopharyngeal carcinoma cell lines in vitro and its mechanism.METHODS: The inhibitory effect on proliferation was evaluated by MTT assay. The radiosensitivity of the cells was compared by colony formation assay. The cell cycle was analyzed by flow cytometry. RESULTS: The maximum non-cytotoxic doses of Tet for CNE1 and CNE2 cells were 1.5 and 1.8 μmol/L, respectively. Compared with radiation group, the cell proliferation in Tet plus radiation group was significantly inhibited on the 4th to 6th days (P<0.01). The mean lethal doses for CNE1 and CNE2 cells in radiation group were (1.26±0.02) Gy and (2.27±0.04) Gy, respectively,and the values changed to (0.73±0.05) Gy and (1.61±0.08) Gy in Tet plus radiation group, respectively, resulting in the sensitivity enhancement ratio of 1.73 and 1.40, respectively (P<0.05). The CNE1 and CNE2 cells in G₂ phase of the cell cycle in radiation group were (42.62±2.07)% and (34.82±2.74)%, respectively, while those in Tet plus radiation group were (17.02±1.87)% and (19.64±4.82)%, respectively. CONCLUSION: Tetrandrine enhances the radiosensitivity of human nasopharyngeal carcinoma cell lines and the mechanism may be related to the abrogation of radiation-induced G₂ phase arrest.