Literature analysis of lung toxicity related to cyclin-dependent kinase 4/6 inhibitors / 中国药房

OBJECTIVE To provide reference for rational use of cyclin-dependent kinase 4/6 （CDK4/6） inhibitors. METHODS Retrieved from Web of Science， PubMed， SpringerLink， CNKI， Wanfang Data and VIP database， and so on， the literature about lung toxicity related to CDK4/6 inhibitors were collected and analyzed statistically with Excel 2013 software. RESULTS A total of 12 literature which met the inclusion and exclusion criteria were included； 13 patients were involved， among which 3 cases were from the United States， 3 from Japan， 2 from India， and 1 from Israel， Spain， France， Australia and Saudi Arabia respectively； all patients were female， aged between 43-89 years， of whom 8 were treated with palbocicilib， 3 with abemacilib， and 2 with ribociclib. The lung toxicity of patients after medication occurred from 1 week to 15 months； the majority of patients were hospitalized with the symptom such as difficulty breathing， chest tightness， shortness of breath， dry cough， etc. The lung toxicity mainly manifested as interstitial lung disease， eosinophilic pneumonia， mediastinal and pulmonary granulomatous reaction， drug-induced pneumonia， diffuse alveolar damage， organizing pneumonia and so on. The shortest treatment duration was 3 weeks， and the longest was 6 months. The treatment measures included drug withdrawal， intravenous use of antibiotics， intravenous use of systemic steroids， oxygen inhalation， and so on； after treatment， 8 patients improved or recovered， and 5 patients died due to deterioration. One patient developed lung toxicity again after reuse of such drugs and must stop drugs permanently. CONCLUSIONS Lung toxicity related to CDK4/6 inhibitors possibly cause mortality. It is necessary to make early judgment， stop the drug in time， and give patients systemic steroids， oxygen inhalation and other treatment measures as soon as possible.

Meta-analysis of efficacy and safety of CDK4/6 inhibitor combined with endocrine in the treatment of HR+/HER2－ breast cancer / 中国药房

OBJECTIVE To evaluate the efficacy and safety of four cyclin-dependent kinase 4/6 （CDK4/6） inhibitors （dalpicilib， abemacilib， ribocilib， palbocilib） combined with endocrine drugs in the treatment of hormone receptor-positive/human epidermal growth factor receptor 2-negative （HR+/HER2－） breast cancer. METHODS Computer searches were conducted on PubMed， the Cochrane Library， Web of Science， Embase， CNKI， Wanfang data and VIP to collect randomized controlled trials （RCTs） about CDK4/6 inhibitors combined with endocrine drugs （trial group） versus endocrine drugs alone or combined with placebo （control group）. The search period was from the establishment of the database to April 2023. After literature screening， data extraction and quality evaluation， a meta-analysis was conducted by using RevMan 5.4.1 software. RESULTS A total of 22 articles were included， involving 15 RCTs with a total of 18 574 patients. The meta-analysis results showed that the progression free survival [HR＝0.77， 95%CI （0.74， 0.79）， P＜0.000 1]， overall survival [HR＝0.91， 95%CI （0.87， 0.94）， P＜0.000 01]， objective response rate [OR＝1.71， 95%CI （1.51， 1.93）， P＜0.000 01] and clinical benefit rate [OR＝1.73， 95%CI （1.52， 1.95）， P＜0.000 01] of the trial group were significantly better than control group. The incidence of adverse drug reactions≥3 levels [OR＝10.28，95%CI （6.97，15.17），P＜0.000 01]， neutropenia [OR＝65.09， 95%CI （36.43， 116.31）， P＜0.000 01]， leukopenia [OR＝22.90， 95%CI （15.40， 34.04）， P＜0.000 01]， anemia [OR＝5.71， 95%CI （4.51， 7.22）， P＜0.000 01]， diarrhea [OR＝ 3.00， 95%CI （1.19， 7.51）， P＜0.05] and nausea [OR＝1.99， 95%CI （1.52， 2.60）， P＜0.000 01] in the trial group was significantly higher than control group. CONCLUSIONS The combination of CDK4/6 inhibitors and endocrine drugs has a significant effect on HR+/HER2－ breast cancer， with a high incidence of adverse reactions， especially hematotoxicity.

Using multiple-fragment amplification combined with Gibson assembly to clone genes with site-directed mutations / 生物工程学报

In order to develop a simple and efficient site-directed mutagenesis solution, the Gibson assembly technique was used to clone the cyclin dependent kinase 4 gene with single or double site mutations, with the aim to simplify the overlap extension PCR. The gene fragments containing site mutations were amplified using a strategy similar to overlap extension PCR. Meanwhile, an empty plasmid was digested by double restriction endonucleases to generate a linearized vector with a short adaptor overlapping with the targeted gene fragments. The gene fragments were directly spliced with the linearized vector by Gibson assembly in an isothermal, single-reaction, creating a recombinant plasmid. After the recombinant plasmids were transformed into competent Escherichia coli DH5α, several clones were screened from each group. Through restriction analysis and DNA sequencing, it was found that the randomly selected clones were 100% target mutants. Since there was neither tedious multiple-round PCR amplification nor frequent DNA extraction operation, and there was no need to digest the original plasmid, this protocol circumvents many factors that may interfere with the conventional site-directed mutagenesis. Hence, genes with single or multiple mutations could be cloned easily and efficiently. In summary, the major defects associated with overlap extension PCR and rolling circle amplification were circumvented in this protocol, making it a good solution for site-directed mutagenesis.

Consensus recommendations for the clinical application of CDK4/6 inhibitors in patients with hormone receptor positive, human epidermal growth factor receptor 2 negative advanced breast cancer / 中华肿瘤杂志

The introduction of cyclin-dependent kinase (CDK) 4/6 inhibitors has revolutionized the clinical management paradigm of hormone receptor (HR) positive/human epidermal growth factor receptor (HER) 2 negative breast cancer. As of today, CDK 4/6 inhibitors including Palbociclib, Ribociclib, and Abemaciclib have been widely approved by regulatory agencies. Randomized clinical trials demonstrated that CDK 4/6 inhibitors in combination with an aromatase inhibitor (AI) or fulvestrant in the first-, second- or later-line setting for HR positive/HER2 negative locally advanced or metastatic breast cancer led to substantial reduction in the risk of disease progression or death. Adverse effects of treatment were manageable and as or better than expected in terms of patient satisfaction. Considering CDK4/6 inhibitors in combination with endocrine therapy being a novel approach in China clinical practice, the panel developed the consensus comprehensively describing the pharmacology properties, monitoring strategy during treatment and adverse events management, to facilitate greater understanding in Chinese oncologists of a whole new therapeutic class of drug, promote accuracy of clinical decision and help reach the ultimate goal of improving survival and quality of life of the target patient population.

Palbociclib in breast cancer neoadjuvant setting

Cyclin-dependent kinase 4/6 inhibitors represent a major advance in breast cancer treatment, emerging as the standard of care of the initial treatment of hormone receptor-positive and HER2-negative metastatic breast cancer. Their activity in this subset of patients leads to interest in their use in the adjuvant and neoadjuvant settings. This case report presents a real-life case of cyclin-dependent kinase 4/6 inhibitors use in a patient initially considered to have stage IV luminal HER2-negative breast cancer with liver metastasis. The discrepancy of treatment response between the breast tumor and liver node led to a repetition of the liver biopsy, which revealed metastasis of a neuroendocrine tumor of unknown primary. The breast tumor showed a partial response, and the initial therapeutic strategy was then redefined for curative intent. While cyclin-dependent kinase 4/6 inhibitors are not yet approved for clinical practice in the neo / adjuvant treatment of hormone receptor-positive breast cancer, this case report portrays a successful example of its application in a neoadjuvant setting.

Research progress of CDK4/6 inhibitors in ER-positive and HER2-negative advanced breast cancer / 国际外科学杂志

Endocrine therapy is one of the standard treatment options for breast cancer which plays an important role in treating patients with estrogen receptor (ER) positive and human epidermal growth factor receptor 2 (HER2) negative breast cancer.However,some patients develop resistance during therapy due to factors such as tumor heterogeneity,which is particularly acute in the treatment of advanced breast cancer.Based on aiming at a rational and effective treatment,some clinical trials recently have demonstrated that compared to endocrine therapy alone,cyclin-dependent kinase 4/6 (CDK4/6) inhibitors combined with endocrine therapy can significantly improve the prognosis of ER-positive,HER2-negative advanced breast cancer.Its main products are Palbociclib,Ribociclib and Abemaciclib.This review mainly focuses on the mechanism and related clinical trials of CDK4/6 inhibitor inhibitors in ER-positive,HER2-negative advanced breast cancer.

Progress of cyclin dependent kinase 4/6 inhibitors in treatment of advanced estrogen receptor-positive/human epidermal growth factor receptor 2-negative breast cancer / 肿瘤研究与临床

Cell cycle disorder is a marker of cancer and has become a key point in the mechanism of endocrine resistance in advanced estrogen receptor-positive/human epidermal growth factor receptor 2-negative (ER+/HER-2-) breast cancer. The combination of cyclin dependent kinase (CDK) 4/6 inhibitors and endocrine therapy in treatment of ER+/HER-2- breast cancer has experienced rapid development and has been commercialized in many countries. This review will describe the mechanisms of action of CDK4/6 inhibitors and their development in advanced ER+/HER-2- breast cancer.

Effect of microRNA-206 on the growth of prostate cancer cells by interfering with the expression of CDK4 and GAK / 国际肿瘤学杂志

Objective To investigate the effect of microRN-206 (miR-206) on the expression of Cyclin-dependent kinase 4 (CDK4) and Cyclin G-associated protein kinase (GAK), and the growth of prostate cancer cells.Methods Prostate cancer cell lines DU-145 and PC-3 were transfected with miR-NC (the control group) or miR-206 (the experimental group).The expressions of CDK4 and GAK mRNA were detected by real-time quantitative PCR (qRT-PCR).The expressions of CDK4 and GAK protein were detected by Western blotting.Cell cycle distribution was detected by flow cytometry.EdU proliferation assay and colony forming assay were used to analyze the cell proliferation ability.Results In DU-145 and PC-3 cells, the expressions of CDK4 mRNA in miR-NC group were 1.00±0.09, 1.00±0.10, the expressions of GAK mRNA were 1.00±0.05, 1.00±0.06.The expressions of CDK4 mRNA in miR-206 group were significantly decreased in DU-145 (0.36±0.18;t=6.572, P=0.001) and PC-3 cell lines (0.43±0.17;t=5.794, P=0.001).The expressions of GAK mRNA were also significantly decreased in DU-145 (0.23±0.04;t=22.420, P<0.001) and PC-3 cell lines (0.32±0.08;t=14.500, P<0.001).Western blotting results were consistent with qRT-PCR results.The results of flow cytometry showed that compared with the miR-NC group of DU-145 and PC-3 cell lines, the percentage of cells in S phase (23.60%±5.68% vs.32.53%±4.52%, t=2.462, P=0.049;22.09%±4.35% vs.30.96%±4.86%, t=2.720, P=0.035) and G2-M phase (16.28%±7.12% vs.26.63%±4.33%, t=2.484, P=0.048;14.60%±1.62% vs.24.68%±7.13%, t=2.758, P=0.033) decreased after transfection of miR-206, and the percentage of cells in G0-G1 phase (60.13%±5.82% vs.40.84%±5.37%, t=4.872, P=0.003;63.31%±3.27% vs.44.36%±3.82%, t=7.533, P<0.001) increased.The results of EdU proliferation assay showed that the proliferation abilities were significantly attenuated after transfection of miR-206 (22.56±3.81 vs.38.90±8.51, t=3.503, P=0.013;25.12±6.42 vs.48.45±8.92, t=4.244, P=0.005).The results of colony formation experiments showed that the numbers of colonies formed by DU-145 and PC-3 in miR-NC group were 218.66±44.59 and 177.35±24.49, respectively.The numbers of colonies formed in miR-206 group were 125.38±32.80 (t=3.370, P=0.015) and 82.65±14.05 (t=6.708, P=0.001), suggesting that cell proliferation ability in miR-206 group was reduced.Conclusion miR-206 significantly inhibits the growth of prostate cancer cells by interfering with the expressions of CDK4 and GAK, suggesting that miR-206 may be a molecular targeted therapy tool for prostate cancer.

Effects of sodium arsenite on Cyclin D1 and Cyclin dependent kinase 4 of human normal liver cells / 中华地方病学杂志

Objective To investigate the effects of sodium arsenite (NaAsO2) on the expressions of Cyclin D1 and Cyclin dependent kinase 4 (CDK4) in human normal liver cells (L-02).Methods L-02 cells were exposed to different doses of NaAsO2 (0,50,100,150 μmol/L) for 24 h,flow cytometry was used to detect the cell cycle,real time quantitative PCR and Western blotting were used to detect the Cyclin D1,CDK4 mRNA and protein expression.Results Cell cycle detection:compared with the control group G0-G1 phase [(60.33 ± 0.40)％],except 50 μmol/L NaAsO2 group [(54.58 ± 0.40)％],the numbers of cells in 100 and 150 μmol/L NaAsO2 groups [(64.60 ± 0.62)％,(83.13 ± 0.25)％] were increased,the differences were statistically significant (all P ＜ 0.05);cell proportion of S phase in the control group,50,100 and 150 μmol/L NaAsO2 groups [(34.35 ± 0.30)％,(29.89 ± 0.32)％,(29.50 ± 0.44)％,(11.93 ± 0.12)％] were decreased,the differences were statistically significant (all P ＜ 0.05);cell proportion of G2-M phase was compared between the control group,50,100 and 150 μmol/L NaAsO2 groups [(5.32 ± 0.11)％,(15.54 ± 0.14)％,(5.90 ± 0.20)％,(4.93 ± 0.15)％],the difference was statistically significant (F =908.359,P ＜ 0.05).Cyclin D1 and CDK4 detection:the expressions of Cyclin D1 mRNA (0.48 ± 0.17,1.00 ± 0.31,1.00 ± 0.21,2.06 ± 0.53) and protein (0.65 ± 0.02,0.64 ± 0.05,0.71 ± 0.10,0.84 ± 0.05) were compared betwee the control group,50,100 and 150 μmol/L NaAsO2 groups,the differences were statistically significant (F =167.886,46.575,all P ＜ 0.05);the expressions of CDK4 mRNA (1.04 ± 0.19,1.00 ± 0.21,1.29 ± 0.22,2.31 ± 0.31) and protein (0.67 ± 0.04,0.74 ± 0.03,0.83 ± 0.07,0.94 ± 0.04) were compared betwee the control group,50,100 and 150 μ mol/L NaAsO2 groups,the differences were statistically significant (F =95.171,145.848,all P ＜ 0.05).Conclusions Treatment of L-02 cells with NaAsO2 has changed the expressions of Cyclin D1,CDK4 mRNA and protein,which leads to L-02 cell cycle arrested at G0-G1 phase,ultimately leads to cell damage.

Effects of a small interfering RNA targeting CDK4 gene on the biological functions of endometrial cancer cells / 中国癌症杂志

Background and purpose: Cyclin-dependent kinase 4 (CDK4) is a kind of protein kinases regulating the cell cycle progression, which has been reported to be overexpressed in endometrial carcinoma tissues. But the role of CDK4 in endometrial carcinogenesis and relative mechanisms has not been identiifed yet. In this study, we used a small interfering RNA targeting CDK4, and explored the effects of CDK4 on endometrial cancer cells HEC-1B biological function and relative mechanisms.Methods:The chemically synthesized small interfering RNA targeting CDK4 (si-CDK4) was transiently transfected into HEC-1B cells;the quantitative real time-PCR assays and Western blot assays were performed to explore the mRNA and protein expression levels of CDK4 and its downstream genes, Rb and p-Rb, in HEC-1B cells upon transfection;Moreover, the CCK-8, lfow cytometry (FCM) and invasion assays were performed to indentify the effects of si-CDK4 on the proliferation, cell cycle distribution, apoptosis and invasion abilities of HEC-1B cells, respectively. Results:The results showed that the mRNA and protein expressions of CDK4 were suppressed in HEC-1B cells upon transfection with si-CDK4 (P<0.01);Suppression of CDK4 inhibited cell proliferation and invasion of HEC-1B cells;the number of cells migrating through the transwell membrane in si-CDK4 group was 117±21, which was much fewer than the cells in si-control (269±39) and untreated groups (262±35) (P<0.01);the early apoptosis rate of cells treated with si-CDK4 [(21.7±3.5)%] was much higher than the untreated [(12.4±2.1)%] and si-control groups [(11.8±1.9)%] (P<0.01);moreover, suppression of CDK4 increased cells in G1 phase (P<0.01) and correspondingly decreased cells in S phase (P<0.01);further Western blot results showed that suppression of CDK4 down-regulated the expression of p-Rb in cells, but did not inlfuence the expression of total Rb. Conclusion:CDK4-siRNA speciifcally and efifciently blocks the constitutively activated CDK4 in human endometrial cancer cells HEC-1B, resulting in tumor suppression.

Functional Inactivation of pRb Associated with Cyclin D1- and Cyclin-dependent Kinase 4 Overexpression Plays A Key Role in Human Pituitary Tumorigenesis

BACKGROUND: Human pituitary adenoma (PA) is a common intracranial tumor, but the mechanism underlying tumorigenesis has not been established. Functional inactivation of retinoblastoma protein (pRb) following cyclin D1- and cyclin-dependent kinase (CDK) 4-dependent hyperphosphorylation is one of the most important mechanisms in tumor cell proliferation. We evaluated immunohistochemical expressions of cyclin D1, CDK4 and phosphorylated pRb (p-pRb) in 50 PAs to investigate a role for functional inactivation of pRb associated with cyclin D1/CDK4 overexpression in pituitary tumorigenesis and to correlate it with clinicopathologic variables. METHODS: Fifty human PAs were immunohistochemically stained for cyclin D1, CDK4 and p-pRb (Thr 356). Correlations between their expression and the clinicopathologic characteristics were statistically analyzed. RESULTS: Cyclin D1 and CDK4 were overexpressed in 56% and 64%, respectively; pRb was hyperphosphorylated in 64%. Forty one cases (82%) showed one or more of these altered expressions. Overexpressions of cyclin D1 and CDK4 were correlated with functional pRb inactivation. Cyclin D1 overexpression was associated with apoplexy and growth hormone production. CONCLUSIONS: Functional inactivation of pRb associated with the cyclin D1/CDK4 overexpression might play a key role in human pituitary tumorigenesis. CDK4 worked in concert with cyclin D1 to hyperphosphorylate pRb. Pituitary apoplexy appeared to be associated with cyclin D1 overexpression.

Expression of CDK4 and cyclin D1 proteins in gastric carcinoma / 上海第二医科大学学报

Objective To investigate the expression and significance of CDK4 and cyclin D1 in gastric carcinoma,and to discuss their correlation with WHO histological classification,TNM stages and lymph node metastasis.Methods Forty paraffin wax specimens from patients with gastric carcinoma were collected and expression of CDK4 and cyclin D1 proteins were detected by immunohistochemistry.Results Immunohistochemistry demonstrated that the positive rates for CDK4 and cyclin D1 protein were 65% and 55%,respectively.There was positive correlation between CDK4 and cyclin D1 proteins(P0.05).Conclusion CDK4 and cyclin D1 proteins are frequently overexpressed in gastric carcinoma and there is a positive relationship between the two proteins,which may be involved in the development of gastric carcinoma.

Expression and Effect of Cyclin Dependent Kinase-4 and p21 in Premature Rats with Hyperoxia-Induced Chronic Lung Disease / 实用儿科临床杂志

Objective To investigate the dynamic change of expression of cyclin dependent kinase-4(CDK4)and cyclin dependent kinase inhibitor(p21)in premature rats with hyperoxia-induced chronic lung disease(CLD)and its role.Methods Eighty premature rats were randomly and equally divided into model group(hyperoxia group)and control group(room air group).CLD was induced by hypemxia exposure.The expression of CDK4 and p21 were observed with immunohistochemical method,and the levels of type I collagen were detected by enzyme-linked immunosorbent assay on days 1,3,7,14 and 21.Results Compared with control group,in model group,the expression of CDK4 protein and level of type I collagen statistically increased,but expression of p21 protein decreased significantly on days 14 and 21.The expression of CDK4 was positive correlated with the degree of fibrosis,and expression of p21 was negative correlated with the degree of fibrosis in model group.Conclusion The expression of CDK4 increases and expression of p21 decreases in premature rats exposed to hyperoxia,which may play an important role in the lung fibrosis.

Grape seed procyanidin extract arrests cell cycle of human bladder cancer cell line BIU87 / 第三军医大学学报

Objective To study the effect of grape seed procyanidin extract(GSPE) on the cell cycle arrest in human bladder cancer BIU87 cells and investigate the underlying molecular mechanism.Methods BIU87 cells were treated with different concentrations of GSPE(50,100 and 200 ?g/ml) and cultured for 24 h.The untreated group served as control.Flow cytometry was used to evaluate the cell cycle,and RT-PCR and Western blotting were used to detect the mRNA and protein expressions of CyclinD1 and cyclin-dependent kinase 4(CDK4).Results GSPE inhibited the cell growth through cell cycle arrest at G1 phrase in a dose-dependent manner.Semi-quantitated RT-PCR and Western blot analyses indicated that GSPE decreased CyclinD1 and CDK4 expressions in a dose-dependent fashion(P

Expression of Cyclin D1 and CDK4 in DMBA-Induced Rat Ovarian Cancer / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Ovarian cancer is a common gynecologic malignancy and the leading cause of death in women. It is typically not diagnosed until it has reached the advanced stages. We performed this study to investigate the roles of the proteins related to the G1 cell cycle in ovarian carcinogenesis. MATERIALS AND METHODS: Immunohistochemistry and Western blot were used to analyse the expression of cyclin Dl and CDK4 in 7, 12-dimethylbenzanthracene- induced ovarian cancer in rats. RESULTS: The Cyclin D1 and CDK4 labelling index was significantly higher in the ovarian cancers than in the normal ovarian surface epithelium of rats. There was no difference among the cancer types. In Western blot analyses, the expression of cyclin Dl and CDK4 in the ovarian cancers was higher than that in the normal ovarian surface epithelium. A positive correlation was observed between the expressions of the CDK4 and Cyclin D1. CONCLUSION: The upregulation of cyclin Dl and CDK4 that occurs in DMBA-induced rat ovarian carcinogenesis is likely to be associated with tumor progression. Further studies are needed to investigate the role and function of cyclin Dl and CDK4 in human ovarian cancer.