Therapeutic effect of Rab11 inhibitor cyclin-dependent kinase inhibitor-73 on liver fibrosis and its related mechanisms / 中华肝胆外科杂志

Objective:To investigate the therapeutic effect and potential molecular mechanisms of cyclin-dependent kinase inhibitor-73 (CDKI-73), the Rab11 inhibitor, on liver fibrosis.Methods:Human LX2 cells were divided into four groups: negative control group, transforming growth factor-β (TGF-β) group, CDKI-73 group and TGF-β+ CDKI-73 group. Fifteen 5-week-old female C57 mice with body weight of (18.04±0.62) g were divided into 3 groups with 5 mice in each group: control group (intraperitoneal injection of olive oil + vehicle gavage), carbon tetrachloride (CCl 4) group (intraperitoneal injection of CCl 4 + vehicle gavage) and CCl 4+ CDKI-73 group (intraperitoneal injection of CCl 4+ CDKI-73 gavage). Another 15 5-week-old female C57 mice with body weight of (18.06±0.34) g were divided into 3 groups with 5 mice in each group: sham operation group (Sham), bile duct ligation (BDL) group + vehicle group (BDL+ vehicle gavage) and bile duct ligation+ CDKI-73 group (BDL+ CDKI-73 gavage). The expression of α-smooth muscle actin (α-SMA) and fibronectin(FN)in LX2 cells were analyzed by Western blot. Masson and Sirius red were used to examine the liver fibrosis after CDKI-73 treatment in vivo. Immunohistochemistry (IHC) was utilized to examine the expression of α-SMA in mice liver. Results:Collagen content assessed by Sirius red and Masson staining and α-SMA expression evaluated by IHC were all increased in CCl 4 group compared with control group ( q=38.47, 24.99, 36.79). Moreover, the collagen content and α-SMA expression in CCl 4 + CDKI-73 treatment group were obviously decreased compared with CCl 4 group ( q=24.72, 14.87, 27.50), and the differences were statistically significant (all P<0.001). Compared with Sham group, collagen content and α-SMA expression in bile duct ligation group were increased ( q=28.23, 41.01, 44.16). Furthermore, in BDL group, after treatment with CDKI-73, the collagen content and α-SMA expression were notably decreased ( q=22.88, 34.31 and 33.97, all P<0.001). Consistent with in vivo results, the relative expression levels of α-SMA and FN protein in TGF-β group were higher than those in TGF-β+ CDKI-73 group (α-SMA: 3.71±0.34 vs. 1.28±0.31; FN: 3.21±0.39 vs. 0.83±0.06, all P<0.001). The mRNA relative expression levels of α-SMA and FN in TGF-β group were higher than those in TGF-β+ CDKI-73 group, and the differences were statistically significant ( P<0.001). However, the relative expression of TGF-β receptor Ⅱ protein in CDKI-73 group was higher than those in negative control group (4.68±0.63 vs. 1.00±0.22, P=0.004). The relative expression level of phosphorylated SMAD2 in TGF-β+ CDKI-73 group was lower than those in TGF-β group (1.67±0.24 vs. 3.99±0.44, P<0.001). Transwell assay showed that 0.5 μmol/L CDKI-73 could effectively inhibit the migration of LX2 cells, and the inhibitory ability became stronger with the increase of CDKI-73 concentration. Conclusion:CDKI-73 can inhibit the activation of hepatic stellate cells and liver fibrosis by inhibiting Rab11-dependent TGF-β signaling pathway both in vivo and in vitro.

Estudio observacional de práctica habitual con palbociclib y hormonoterapia para carcinoma de mama avanzado / Observational real world data with palbociclib associated to hormone therapy for advanced breast carcinoma

Objetivo: Los inhibidores de quinasas dependientes de ciclina CDK4y CDK6 poseen efecto sinérgico al asociarse con hormonoterapia. Suuso está extendido en primera y sucesivas líneas de carcinoma de mamaavanzado tipo luminal por mejorar la supervivencia libre de progresión.Los objetivos de nuestro estudio se basaron en analizar la evolución clínica y la toxicidad presentada en las pacientes tratadas en nuestro centrocon palbociclib, así como relacionar la evolución con las diferentes variables clínico-patológicas.Método: El estudio, de tipo observacional y retrospectivo, recogió datosde pacientes con cáncer de mama avanzado o metastásico tratados conhormonoterapia y palbociclib en el Hospital Universitario de Cabueñesentre los años 2017 y 2020. Se analizaron diferentes variables clínicopatológicas, así como información sobre toxicidad y supervivencia.Resultados: Un total de 72 mujeres y 1 varón con una mediana deedad de 63 años recibieron palbociclib asociado a inhibidor de aromatasa o fulvestrant. En primera línea la supervivencia libre de progresión fuede 22 meses, y en segunda o sucesivas líneas de 13 meses. El 95,9% de las pacientes presentaron algún tipo de efecto adverso, principalmentehematológico. No se produjo ningún abandono por toxicidad, aunquelos retrasos y los ajustes de dosis fueron frecuentes (61,7% y 42,7%, respectivamente). Solo la situación funcional al inicio del tratamiento influyóde manera significativa en la supervivencia libre de progresión (22 mesesen ECOG 0 versus 12 meses en ECOG ≥ 1; p = 0,021). (AU)

Objective: Cyclin-dependent kinase 4/6 inhibitors have a synergisticeffect in combination with endocrine therapy. This combination is usedas first and subsequent-line treatment for advanced luminal breast carcinoma because it increases progression-free survival. We analysed clinicalcourse and toxicity in patients treated with palbociclib in our hospital anddetermined potential associations between these variables and clinicopathological variables.Method: Observational retrospective study including patients withadvanced or metastatic breast cancer treated with palbociclib plus endocrine therapy at the Hospital Universitario de Cabueñes between 2017and 2020. We analysed clinicopathological variables, toxicity, and survival.Results: In total, 72 women and 1 man (median age: 63 years) receivedpalbociclib plus an aromatase inhibitor or fulvestrant. When used as firstline treatment, progression-free survival was 22 months, and as secondand subsequent-line treatment, progression-free survival was 13 months.Adverse effects (mainly haematological) were experienced by nearly all patients (95.9%). Treatment was not discontinued because of toxicity inany patient, although delays and dose adjustments were common (61.7%and 42.7%, respectively). Performance status alone had a significantimpact on progression-free survival (22 months in patients with ECOG 0vs 12 months in patients with ECOG ≥ 1; P = 0.021). (AU)

Research progress of radiotherapy and CDK4/6 inhibitors in metastatic breast cancer / 国际肿瘤学杂志

Cyclin-dependent kinase (CDK) 4/6 inhibitors are a new class of molecular targeted drugs, which can enhance radiotherapy sensitivity by anti angiogenesis, inhibiting DNA damage repair and inhibiting mammalian target of rapamycin signal transduction. Existing clinical trials have confirmed that radiotherapy combined with CDK4/6 inhibitors can effectively control the local symptoms of breast cancer metastases and prolong progression-free survival. Compared with CDK4/6 inhibitors alone, the combination with radiotherapy does not significantly increase the incidence and severity of adverse reactions. However, there are also reports about severe adverse reactions of normal tissue happened in the radiation field in individual cases of combined treatment, and its efficacy and safety need to be clarified by more basic and clinical observational researches.

Effects of 12-lipoxygenase and angiotensin Ⅱ on p21, p27 and p57 in rat diabetic glomeruli / 中华肾脏病杂志

Objective To investigate the effects of 12-lipoxygenase (12-LO) and angiotensin Ⅱ (Ang Ⅱ) on the CIP/KIP family of cyclin-dependent kinase inhibitors (CKIs) p21,p27 and p57 related to cell hypertrophy.Methods Mesangial cells were treated with high glucose for 24 hours and 48 hours respectively.12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE] and Ang Ⅱ were infused to rats by osmotic mini-pump for 1 week and 2 weeks respectively.Rats fed high fat diet were received low dose streptozotocin (STZ) to make type 2 diabetes (DN).The rats were divided into normal control group,DN group,DN+Ang Ⅱ type 1 receptor blocker (ARB) group or 12-LO inhibitor (CDC) group.DN+ARB rats were treated by losartan for 6 weeks,and DN+CDC rats were treated for 8 weeks.Urine albumin and protein expressions of p21,p27 and p57 were detected by ELISA and Western blotting respectively.Glomeruli injury and expressions of p21 and p27 were detected by PAS staining and immunohistochemistry respectively.Results High glucose increased p21 and p27 protein expression in mesangial cells significantly compared with the relative control (all P ＜ 0.05),but had no effect on p57.Ang Ⅱ increased p27 protein expression in gloneruli significantly (P ＜ 0.05),but had no effect on p21 and p57 protein expression.12(S)-HETE increased both p21 and p27 protein expression in glomeruli significantly (all P ＜ 0.05),but had no effect on p57 protein expression.Blood glucose,kidney/body weight,urinary protein,and glomerular p21 and p27 protein expressions were increased in DN group (all P ＜ 0.05) compared with those in control group,with little change of p57 protein expression (P ＜ 0.05).Moreover,glomerular hypertrophy and extra cellular matrix accumulation were observed in DN group.However,urine protein,kidney/body weight,renal injury,but not blood glucose,were decreased in DN+ARB group and DN+CDC group compared with DN group respectively (P＜ 0.05).Further DN+CDC rats had decreased both p21 and p27 protein expressions in glomeruli,but DN+ ARB rats only had decreased p27 protein expression (all P ＜ 0.05).Conclusions 12-LO may induce both p21 and p27 protein expression in DN glomeruli,but Ang Ⅱ may induce only p27 expression.

CDK4/6 Inhibition Controls Proliferation of Bladder Cancer and Transcription of RB1.

PURPOSE: The retinoblastoma signaling network is frequently altered in advanced bladder cancer. We investigated the potential of CDK4/6 as a therapeutic target and determined biomarkers for patient stratification. MATERIALS AND METHODS: Genetic alterations were analyzed using public databases, including TCGA (The Cancer Genome Atlas), COSMIC (Catalogue of Somatic Mutations in Cancer) and CCLE (Cancer Cell Line Encyclopedia). Effects of the CDK4/6-inhibitor PD-0332991 or LY2835219 were examined in 10 bladder cancer cell lines by immunoblot, cell viability, apoptosis and cell cycle progression. Efficacy of the PD-0332991 and cisplatin combination was analyzed using the combination index. Gene expression level was determined by quantitative polymerase chain reaction. Cytomegalovirus promoter regulated recombinant retinoblastoma was used for reconstitution. Three-dimensional xenografts were grown on chicken chorioallantoic membrane and analyzed by measuring tumor weight and immunohistochemical expression of total retinoblastoma and Ki-67. RESULTS: PD-0332991 treatment decreased the proliferation of retinoblastoma positive bladder cancer cell lines and was synergistic in combination with cisplatin. PD-0332991 or LY2835219 treatment decreased the phosphorylation, total protein and transcript level of retinoblastoma. Treatment resulted in a decrease in E2F target gene expression (CCNA2 and CCNE2) and cell cycle progression from G0/G1 to the S-phase but did not affect apoptosis. In retinoblastoma negative cells reconstituted with recombinant retinoblastoma PD-0332991 affected only phosphorylation and not the total retinoblastoma level. These cells remained resistant to treatment. In 3-dimensional retinoblastoma xenografts, treatment resulted in reduced tumor weight and decreased expression of total retinoblastoma and Ki-67. CONCLUSIONS: We provide preclinical evidence that CDK4/6 inhibition is a potential therapeutic strategy for retinoblastoma positive bladder cancer that probably acts by negatively regulating retinoblastoma transcription.

Actinic cheilitis and squamous cell carcinoma of the lip: clinical, histopathological and immunogenetic aspects / Queilite actínica e carcinoma espinocelular do lábio: aspectos clínicos, histopatológicos e imunogenéticos

Actinic cheilitis is the main precancerous lesion of the lip. Squamous cell carcinoma of the lip is reported together with oral carcinomas in the Brazilian official statistics. Overall, they account for 40% of the head and neck carcinomas. In general, physicians and dentists know little about what causes oral tumor development and progression. Tumor suppressor genes and cell proliferation regulatory proteins play a role in the progression of actinic cheilitis to squamous cell carcinoma and in its biological behavior. Knowledge on prognostic and diagnostic markers has a positive impact on the follow-up of these patients.

Queilite actínica é a principal lesão pré-neoplásica do lábio. O carcinoma espinocelular do lábio é incluído nas estatísticas brasileiras junto com os cânceres de boca e, em conjunto, somam 40% dos cânceres de cabeça e pescoço. Há certo desconhecimento médico e odontológico em geral quanto aos fatores relacionados à carcinogênese e à progressão de tumores de boca. Genes de supressão tumoral e proteínas regulatórias de proliferação celular exercem papel na evolução da queilite actínica para carcinoma espinocelular e no comportamento biológico deste. O conhecimento de marcadores de diagnóstico e prognóstico e sua investigação têm utilidade no acompanhamento de tais pacientes.