Skip to main content
NCBI home page
Journal of Southern Medical University logoLink to Journal of Southern Medical University
. 2020 Dec 20;40(12):1784–1792. [Article in Chinese] doi: 10.12122/j.issn.1673-4254.2020.12.13

Palbociclib可诱导人肾小管上皮细胞周期阻滞及衰老

Palbociclib induces cell cycle arrest and senescence of human renal tubular epithelial cells in vitro

Liuwei HUANG 1, Yanting SHEN 1, Chongbin LIU 1, Caizhen LI 1, Jun WANG 1,*
PMCID: PMC7835700  PMID: 33380388

Abstract

Objective

To investigate the effect of palbociclib on cell cycle progression and proliferation of human renal tubular epithelial cells.

Methods

Human renal tubular epithelial cell line HK-2 was treated with 1, 5, 10, and 20 μmol/L of palbociclib, and the changes in cell proliferation and viability were examined by cell counting and CCK8 assay. EDU staining was used to assess the proliferation of HK-2 cells following palbiciclib treatment at different concentrations for 5 days. The effect of palbociclib on cell cycle distribution of HK-2 cells was evaluated using flow cytometry. SA-β-Gal staining and C12FDG senescence staining were used to detect senescence phenotypes of HK-2 cells after palbociclib treatment at different concentrations for 5 days. The relative mRNA expression levels of P16, P21, and P53 and the genes associated with senescence-related secretion phenotypes were detected by RT-PCR, and the protein expressions of P16, P21 and P53 were detected by Western blotting.

Results

Palbociclib inhibited HK-2 cell proliferation and induced cell cycle arrest in G1 phase. Compared with the control cells, HK-2 cells treated with high-dose (10 μmol/L) palbociclib exhibited significantly suppressed cell proliferation activity, and the inhibitory effect was the most obvious on day 5 (P < 0.01). Palbociclib treatment significantly reduced the number of cells in S phase (P < 0.01) and induced senescence of HK-2 cells. The results of SA-β-Gal and C12FDG senescence staining showed a significantly enhanced activity of intracellular senescence-related galactosidase in palbociclib-treated HK-2 cells, suggesting significant senescence of the cells (P < 0.01). RT-PCR and Western blotting showed that palbociclib treatment significantly increased the mRNA and protein expression levels of P16, P21, and P53 in HK-2 cells (P < 0.01); the mRNA expression levels of senescence-related secretory factors also increased significantly in HK-2 cells after palbociclib treatment (P < 0.01).

Conclusions

Palbociclib induces HK-2 cell senescence by causing cell growth arrest and delaying cell cycle progression.

Keywords: palbociclib, renal tubule epithelial cells, cell proliferation, cellular senescence

细胞周期蛋白依赖性激酶（CDKs）为调节细胞增殖的关键酶，主要通过调节细胞周期检查点及调控胞内外信号转导通路以促进细胞增殖[1-2]。CDKs（如CDK4和CDK6）失调是肿瘤细胞的特征之一，阻断CDK4/6活性可抑制肿瘤细胞增殖[3-4]。Palbociclib（PD-0332991）是近年来首个获准用于乳腺癌治疗的CDK4/6抑制剂，其作用主要通过阻止cyclin D-CDK4/6磷酸化而导致细胞周期停滞于G1期[5-6]。因此，Palbociclib主要通过抑制肿瘤细胞增殖以发挥抗肿瘤的疗效[7-9]。Palbociclib在诱导细胞生长周期停滞后，可进一步促进细胞发生衰老[10]。在多种疾病的动物模型中均发现衰老细胞可促进器官纤维化进展[11-12]。尽管Palbociclib在Ⅱ期试验中耐受性良好，并且正在Ⅲ期临床研究试验中用于乳腺癌的治疗，但仍可能出现其通过诱导衰老细胞的产生进一步促进机体其它组织和器官纤维化进展的潜在安全隐患。目前并无相关文献报道Palbociclib对肾脏的影响，在合并慢性肾脏病（CKD）的肿瘤患者中应用Palbociclib进行抗肿瘤治疗时，也可能促进CKD患者肾脏纤维化进展。本文拟通过体外实验证实Palbociclib是否可诱导肾小管上皮细胞发生衰老。

1. 材料和方法

1.1. 主要试剂

Palbociclib（PD-0332991，Selleck）；1.0 g DMEM低糖完全培养基、澳洲胎牛血清和胰蛋白酶（Gibco）；CCK8检测试剂盒（Dojindo）；台盼蓝染料（CST）；细胞计数板（Marienfeld）；流式细胞周期检测试剂盒（联科）；EDU检测试剂盒（锐博）；SA-β-Gal染色试剂盒（CST）；C12FDG染料（Life）；Trizol细胞裂解液（Takara）；RNA逆转录试剂盒（Takara）；ChamQ SYBR qPCR Master Mix（诺唯赞）；DAPI荧光抗衰减封片剂（中杉金桥）；BCA蛋白浓度检测试剂盒（碧云天）；10× RIPA细胞裂解液（CST）；兔抗鼠P16单克隆抗体（Santa Cruz）；兔抗鼠P21单克隆抗体（Santa Cruz）；兔抗P53单克隆抗体（CST）；兔抗鼠β-Actin单克隆抗体（Proteintech）；山羊抗鼠荧光二抗（Licor）；山羊抗兔荧光二抗（Licor）。

1.2. 方法

1.2.1. 细胞培养及分组

实验用人肾小管上皮细胞系（HK-2细胞）来自广东医科大学肾脏疾病研究所刘华锋教授团队。HK-2细胞培养于含有10%澳洲胎牛血清（FBS）的低糖DMEM培养基中。细胞于37 ℃，5% CO2恒温孵箱中进行培养，48 h后用胰酶进行消化传代。细胞状态良好且处于生长对数期时，将细胞分组种板，种板数为1.0×105/皿，之后进行药物处理，实验分组为空白对照组和Palbociclib组。将不同浓度的Palbocilib（1、5、10、20 μmol/L）对HK-2细胞作用不同天数（1、3、5、7 d），其中在第0天加入药物工作液后，后分别在第2、4、6天时更换新鲜的药物工作液。根据实验中所得结果，以确定Palbocilib的最佳工作浓度和作用时间，并应用于后续实验中。

1.2.2. 细胞计数

每小皿细胞种板数为1.0×105，每组设置3个小皿，细胞干预结束后，胰酶消化细胞，用台盼蓝染料混合细胞悬液，加入细胞计数板进行计数，待细胞沉降1 min后，于正置光学显微镜10×镜下计数后，求出细胞悬液中的总细胞数。

1.2.3. 细胞活力与增殖分析

采用CCK8法检测细胞活力，每组设置3个复孔。每孔细胞种板数约为1000个，细胞干预结束后，每孔加入10 μL CCK8工作液，避光孵育1~4 h，用酶标仪测定A450 nm值以评估细胞的活力与增殖情况。

1.2.4. EDU染色

采用EDU染色检测细胞增殖情况。细胞分组爬片种板，每小皿细胞种板数为1.0×105，药物干预结束后，弃上清，PBS清洗1次；根据EDU试剂盒相关操作说明，先后分别用试剂A和EDU工作液进行孵育，最后加入甲醇清洗，洗2次，5 min/次；含DAPI封片剂封片，于4 ℃避光保存。

1.2.5. 流式检测细胞周期

细胞分组种板，每小皿细胞种板数为1.0×105，药物干预结束后，收集1×106细胞，加入1 mL DNA Staining solution和10 μL Permeabilization solution, 涡旋震荡5~10 s，混匀。室温避光孵育30 min；选择最低上样速度，于流式细胞仪上检测。细胞周期结果通过Flow Jo7.6.1软件进行分析。

1.2.6. SA-β-Gal染色

采用SA-β-Gal染色检测细胞衰老表型。细胞分组爬片种板，每小皿细胞种板数为1.0×105，药物干预结束后，弃上清，1× Fixative solution对细胞室温固定15 min；按说明书配制β-Gal工作液，用pH测定仪调节pH值至6.0，加入细胞后，于37 ℃无CO2孵箱孵育过夜；移除染色液，PBS洗3次，5 min/次，水性封片剂封片后，4 ℃保存，于Olympus正置显微镜×20和×40进行拍片。

1.2.7. C12FDG染色

采用SA-β-Gal染色检测细胞衰老表型。细胞分组爬片种板，每小皿细胞种板数为1.0×105，药物干预结束后，弃上清，多聚甲醛固定液对细胞室温固定15 min；将C12FDG工作液（25 μmol/L）加入细胞中，于37 ℃ CO2培养箱避光孵育2 h，移除染色液，PBS洗3次，5 min/次，含DAPI的抗荧光衰减剂封片，4 ℃避光保存，于Olympus免疫荧光共聚焦显微镜20×和40×进行拍片。

1.2.8. 实时荧光定量PCR

细胞样本通过Trizol裂解液充分裂解以提取总RNA。紫外可见分光光度仪测定RNA样本浓度及纯度，后逆转录为cDNA。实验涉及相关引物序列详见表 1。所得cDNA样本通过q-RT PCR仪进行扩增，扩增条件为：95 ℃ 5 min预变性，95 ℃ 15 s，60 ℃ 1 min，循环40次，95 ℃ 15 s，60 ℃ 1 min，95 ℃ 15 s，溶解。β-Actin作为校正内参，目的基因相对表达量通过2-ΔΔCt法进行计算。

1.

文中涉及相关引物序列

Sequences of primers for RT-PCR

Gene Primer sequence
Human β-actin
Upstream GATTCCTATGTGGGCGACGA
Downstream AGGTCTCAAACATGATCTGGGT
Human P16
Upstream ATGGAGCCTTCGGCTGACT
Downstream GTAACTATTCGGTGCGTTGGG
Human P21
Upstream GCCTGTCCACCATCTCCCTAT
Downstream TTCAAGCACAGTTATTCTGGACCTT
Human P53
Upstream ACCTATGGAAACTACTTCCTGAAA
Downstream CTGGCATTCTGGGAGCTTCA
Human IL-1β
Upstream CTGTCCTGCGTGTTGAAAGA
Downstream TTGGGTAATTTTTGGGATCTACA
Human IFN-β
Upstream ATGACCAACAAGTGTCTCCTCC
Downstream GGAATCCAAGCAAGTTGTAGCTC
Human IL-6
Upstream ACTCACCTCTTCAGAACGAATTG
Downstream CCATCTTTGGAAGGTTCAGGTTG
Human IL-8
Upstream TCTTCATTGACCAAGGAAATCGG
Downstream TCCGGGGTGCATTATCTCTAC
Open in a new tab

1.2.9. 蛋白质免疫印迹实验

细胞样本加入含有蛋白酶抑制剂的RIPA裂解液进行充分裂解，以收集细胞蛋白质。BCA法测定蛋白质样本浓度。通过聚丙烯酰胺凝胶电泳进行等量蛋白分离后转移至NC膜上，室温下封闭30 min，一抗4 ℃孵育慢摇过夜，荧光二抗室温孵育1 h后于化学发光仪进行条带显影。蛋白条带定量通过Image J软件进行分析。

1.3. 统计学处理

统计软件为SPSS19.0，所有数据均通过均数±标准差表示，组间差异采用独立样本t检验或单因素OneWay ANOVA分析进行比较，P < 0.05认为差异有统计学意义。

2. 结果

2.1. 不同浓度及不同作用时间Palbociclib对肾小管上皮细胞活性的影响

与对照组相比，随着作用时间延长，低中剂量（1、5 μmol/L）Palbociclib组并不会引起HK-2细胞生长状态改变，而高剂量（10 μmol/L）Palbociclib组在第3天时，HK-2细胞进入生长停滞状态，到第5天时细胞总数基本与第3天持平（P < 0.01），且于镜下观察时并未发现细胞飘起，至第7天时，极少数HK-2细胞出现漂浮；更高剂量（20 μmol/L）Palbociclib组在第1天时，少数细胞漂浮，第3天时，约半数以上的细胞漂浮，至第5和7天，细胞全部漂浮，表现出了明显药物毒性（P < 0.01，图 1A）。

1.

1

Open in a new tab

Palbociclib对肾小管上皮细胞生长情况的影响

Effect of palbociclib on growth of HK-2 cells. A: Cell counts after treatment with 0, 1, 5, 10 and 20 μmol/L palbciclib for 0, 1, 3, 5 and 7 days. B: Inhibition of HK-2 cell growth after palbciclib treatment at 0, 1, 5, 10 and 20 μmol/L for 0, 1, 3, 5 and 7 days shown by CCK8 assay. **P < 0.01 vs control group.

与对照组相比，低剂量（1、5 μmol/L）Palbociclib组对HK-2细胞活性影响较小，高剂量（10 μmol/L）Palbociclib组在第3天时，明显抑制HK-2细胞的活性，且随着作用时间延长，HK-2细胞活性仍持续降低（P < 0.01），而更高剂量（20 μmol/L）Palbociclib组在第1天已表现出对HK-2细胞活性的明显影响，到第5天时HK-2细胞已全数死亡（图 1B）。

2.2. Palbociclib引起肾小管上皮细胞增殖抑制

分别用0、1、5、10 μmol/L的Palbociclib处理后，在第5天时进一步通过EDU染色检测HK-2细胞的增殖情况。相比于对照组来说，低中剂量（1、5 μmol/L）的Palbociclib对HK-2细胞的细胞增殖无明显影响，高剂量（10 μmol/L）Palbociclib组中，HK-2细胞增殖抑制效果明显（P < 0.01，图 2A、B）。

2.

2

Open in a new tab

Palbociclib对肾小管上皮细胞增殖情况的影响

Effect of palbociclib on proliferation of HK-2 cells. A: EDU staining of palbociclib-treated HK-2 cells at the doses of 0, 1, 5 and 10 μmol/L for 5 days (scale bar=50 μm). B: Percentage of palbocilib-treated HK-2 cells positive for EDU staining. *P < 0.05, **P < 0.01 vs control group.

2.3. Palbociclib致肾小管上皮细胞生长周期停滞于G1-S期

与对照组相比，Palbociclib处理组的HK-2细胞中，处于G0/G1期的细胞百分比明显增加，进入S期的细胞数也明显减少（P < 0.01，图 3A、B）。

3.

3

Open in a new tab

Palbociclib对肾小管上皮细胞生长周期的影响

Effect of palbociclib on cell cycle distribution of HK-2 cells. A: Distribution of cell cycle in HK-2 cells after treatment with 10 μmol/L palbociclib for 5 days. B: Accumulated percentages of HK-2 cells in G1 phase after treatment with 10 μmol/L palbociclib for 5 days. *P < 0.05 vs control group.

2.4. Palbociclib可上调肾小管上皮细胞衰老标志物表达

我们将不同浓度（0、1、5、10 μmol/L）的Palbociclib作用于HK-2细胞5 d后，行SA-β-Gal染色检测HK-2细胞是否发生衰老。SA-β-gal染色阳性细胞通常表现为细胞体积变大，且胞质中存在高活性的β-半乳糖苷酶。与对照组相比，低剂量（1、5 μmol/L）Palbociclib作用的HK-2细胞表现为SA-β-gal染色阴性，而高剂量（10 μmol/L）Palbociclib作用下HK-2细胞的胞质中可检测到大量蓝色络合物沉积，表现出SA-β-gal染色强阳性（P < 0.01，图 4A、C）。应用免疫荧光检测SA-β-gal的作用底物C12FDG判断Palbociclib诱导的HK-2细胞衰老发生情况，所得结果与SA-β-gal染色一致（图 4B、D）。

4.

4

Open in a new tab

Palbociclib可诱导肾小管上皮细胞衰老

Palbociclib induces senescence of HK-2 cells. A: SA β-gal staining of palbociclib-treated HK-2 cells at the doses of 0, 1, 5 and 10 μmol/L for 5 days, which exhibited extended shape, enlarged cell nuclei, and positive SA-β-gal staining in the cytoplasm (scale bar=100 μm). B: C12FDG staining of palbociclib-treated HK-2 cells (scale bar=50 μm). C: Percentage of palbocilib-treated HK-2 cells positive for SA-β-Gal staining. D: Percentage of palbocilib-treated HK-2 cells positive for C12FDG staining. *P < 0.05, **P < 0.01 vs control group.

与对照组相比，Palbociclib作用的HK-2细胞中P16、P21和P53的mRNA表达水平显著升高（P < 0.01，图 5A~C）。免疫印迹技术检测结果表明，Palbociclib也可上调HK-2细胞中P16、P21和P53的蛋白表达水平（P < 0.01，图 5D~H）。

5.

5

Open in a new tab

Palbociclib上调肾小管上皮细胞细胞周期因子mRNA及蛋白表达

Expressions of P16, P21 and P53 in HK-2 cells exposed to 10 μmol/L palbociclib for 5 days. A-C: RT-PCR for detecting P16, P21 and P53 mRNA expressions in HK-2 cells after the treatment. D: Western blotting for detecting P16 protein expression in palbociclib-treated HK-2 cells. E: Quantitative analysis of the level of P16 proteins in palbociclib-treated HK-2 cells. F: Western blotting for detecting P21 and P53 protein expressions in palbociclib-treated HK-2 cells. G-H: Quantitative analysis of the level of P21 and P53 proteins in palbociclib-treated HK-2 cells. *P < 0.05, **P < 0.01 vs control group.

2.5. Palbociclib促进肾小管上皮细胞合成衰老相关分泌表型因子

衰老细胞所分泌的细胞因子、趋化因子及生长因子等则被称为衰老相关的分泌表型（SASP）。与对照组相比，Palbociclib可诱导HK-2细胞IFN、IL-1、IL-6和IL-8等mRNA表达升高（P < 0.01，图 6）。

6.

6

Open in a new tab

Palbociclib上调肾小管上皮细胞因子mRNA表达衰老相关分泌表型

Palbociclib promotes expression of senescence-associated secretory phenotypes (SASP) in HK-2 cells. A-D: RT-PCR for detecting IFN-β and SASP (IL-1β, IL-6 and IL-8) mRNA expressions in HK-2 cells exposed to 10 μmol/L palbociclib for 5 days. **P < 0.01 vs control group.

3. 讨论

肿瘤与CKD间存在密切关联，一方面慢性肾脏病患者肿瘤发生率较正常人群增高，另一方面肿瘤患者中CKD发病率较正常人群也明显升高[13-14]。在导致肿瘤患者CKD发病增高的机制中抗肿瘤药物的作用日益引起关注[15]。

Palbociclib作为一种新型的抗肿瘤药物，其属于CDK4/6周期蛋白抑制剂，临床上主要用于乳腺癌的治疗[16-17]。Palbociclib主要通过诱导肿瘤细胞生长周期停滞而起到抗肿瘤的作用[8, 16]。本研究在体外实验中，将Palbociclib作用于HK-2细胞可明显观察到细胞生长周期停滞，并诱导细胞进入衰老状态。我们在本实验中分别采用了不同浓度Palbociclib作用于HK-2细胞，结果表明，当Palbociclib的工作浓度达到10 μmol/L时，即可对HK-2细胞表现出明显的增殖抑制作用，并诱导HK-2细胞衰老。除了衰老表型的相关检测，我们进一步检测了P16、P21和P53等细胞衰老标志物，发现Palbociclib作用下HK-2细胞内衰老标志蛋白表达明显升高，同时可使得HK-2细胞合成衰老分泌相关表型因子增加，这说明，Palbociclib可诱导肾小管上皮细胞发生衰老。

衰老细胞处于一种稳定的生长停滞状态，其特征在于细胞生长周期不可逆性阻滞[18]。细胞周期蛋白等调控分子对细胞周期中G1-S期和G2-M期的进程至关重要，P16、P21等蛋白可通过结合细胞周期蛋白进而调控细胞衰老[19]。P53/P21WAFI和Rb-P16INK4A通路目前被认为是调控细胞衰老的重要机制。最近研究指出，启动P53/P21WAFI途径可触发细胞衰老反应，随后激活RbP16INK4A通路以维持细胞衰老状态[20-21]。CDK4/6周期蛋白抑制剂作用于肿瘤细胞后，P16表达上调，此外，过表达P16可促进细胞发生衰老[22-24]。本研究中，Palbociclib作用于肾小管上皮细胞后，流式细胞仪结果分析表明细胞周期明显停滞于G1期，随着作用时间延长，肾小管上皮细胞中的SA-β-Gal酶活性明显升高，并且，P53、P21及P16等细胞周期相关调控分子表达明显上调，这说明Palbociclib诱导肾小管上皮细胞衰老可能依赖于P53/P21WAFI或Rb-P16INK4A途径。

衰老细胞可分泌大量衰老相关分泌表型（SASP），进而引发免疫应答反应[25]。一方面，SASP可以诱导免疫细胞募集，介导肿瘤清除[26]；另一方面，SASP可以创造促肿瘤的微环境，进一步促进旁分泌性衰老[27]。此外，SASP还可诱导细胞重塑[28]，未发生衰老的肿瘤细胞转化成癌症干细胞后更具侵袭性[29]。因此，CDK4/6抑制剂诱导产生的衰老细胞，若持久性存在则可能对短期治疗以及癌症转移和复发产生潜在的影响。本研究中，Palbociclib作用下可促进肾小管上皮细胞发生衰老并合成SASP，这提示Palbociclib在临床中应用于抗肿瘤的同时，可能会对患者肾功能的远期预后产生影响。近期研究提出，细胞衰老是促进慢性退行性疾病进展的重要机制[30-32]。衰老细胞通过分泌SASP因子引发慢性炎症反应及纤维化，这是衰老导致器官纤维化的主要机制[33-35]。既往研究显示急性肾损伤所致的肾小管上皮细胞衰老可引发肾脏炎症反应及肾脏纤维化[36-38]。我们之前的研究表明，在顺铂致小鼠肾脏纤维化模型中顺铂所致肾小管上皮细胞衰老发挥了重要作用[39]。因而肾脏衰老细胞被认为是引发肾脏纤维化的重要因素。肿瘤患者如长期应用Palbociclib也可能通过诱发肾脏细胞衰老而促进肾脏纤维化。

综上所述，本研究发现Palbociclib可抑制肾小管上皮细胞增殖，并诱导肾小管上皮细胞衰老。这提示我们在临床中对肿瘤患者尤其是合并有CKD的肿瘤患者应用Palbociclib时，应注意患者肾脏纤维化的进展及远期肾功能的衰退。

Biography

黄柳维，硕士，E-mail: 1027300156@qq.com

Funding Statement

国家自然科学基金（81470972）

Supported by National Natural Science Foundation of China (81470972)

Contributor Information

黄 柳维 (Liuwei HUANG), Email: 1027300156@qq.com.

王 骏 (Jun WANG), Email: wangjun1975@smu.edu.cn.

References

  • 1.Cao LH, Chen F, Yang XM, et al. Phylogenetic analysis of CDK and cyclin proteins in premetazoan lineages. BMC Evol Biol. 2014;14:10. doi: 10.1186/1471-2148-14-10. [Cao LH, Chen F, Yang XM, et al. Phylogenetic analysis of CDK and cyclin proteins in premetazoan lineages[J]. BMC Evol Biol, 2014, 14: 10.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2.Fouty BW, Grimison B, Fagan KA, et al. P27(Kip1) is important in modulating pulmonary artery smooth muscle cell proliferation. Am J Respir Cell Mol Biol. 2001;25(5):652–8. doi: 10.1165/ajrcmb.25.5.4592. [Fouty BW, Grimison B, Fagan KA, et al. P27(Kip1) is important in modulating pulmonary artery smooth muscle cell proliferation[J]. Am J Respir Cell Mol Biol, 2001, 25(5): 652-8.] [DOI] [PubMed] [Google Scholar]
  • 3.Spring LM, Wander SA, Andre F, et al. Cyclin-dependent kinase 4 and 6 inhibitors for hormone receptor-positive breast cancer: past, present, and future. Lancet. 2020;395(10226):817–27. doi: 10.1016/S0140-6736(20)30165-3. [Spring LM, Wander SA, Andre F, et al. Cyclin-dependent kinase 4 and 6 inhibitors for hormone receptor-positive breast cancer: past, present, and future[J]. Lancet, 2020, 395(10226): 817-27.] [DOI] [PubMed] [Google Scholar]
  • 4.Malumbres M, Barbacid M. Cell cycle, CDKS and cancer: a changing paradigm. Nat Rev Cancer. 2009;9(3):153–66. doi: 10.1038/nrc2602. [Malumbres M, Barbacid M. Cell cycle, CDKS and cancer: a changing paradigm[J]. Nat Rev Cancer, 2009, 9(3): 153-66.] [DOI] [PubMed] [Google Scholar]
  • 5.Clark AS, Karasic TB, DeMichele A, et al. Palbociclib (PD0332991)-a selective and potent cyclin-dependent kinase inhibitor: a review of pharmacodynamics and clinical development. JAMA Oncol. 2016;2(2):253–60. doi: 10.1001/jamaoncol.2015.4701. [Clark AS, Karasic TB, DeMichele A, et al. Palbociclib (PD0332991)-a selective and potent cyclin-dependent kinase inhibitor: a review of pharmacodynamics and clinical development [J]. JAMA Oncol, 2016, 2(2): 253-60.] [DOI] [PubMed] [Google Scholar]
  • 6.Ouyang ZX, Wang SS, Zeng M, et al. Therapeutic effect of palbociclib in chondrosarcoma: implication of cyclin-dependent kinase 4 as a potential target. Cell Commun Signal. 2019;17(1):17. doi: 10.1186/s12964-019-0327-5. [Ouyang ZX, Wang SS, Zeng M, et al. Therapeutic effect of palbociclib in chondrosarcoma: implication of cyclin-dependent kinase 4 as a potential target[J]. Cell Commun Signal, 2019, 17(1): 17.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Finn RS, Crown JP, Lang I, et al. The cyclin-dependent kinase 4/6 inhibitor palbociclib in combination with letrozole versus letrozole alone as first-line treatment of oestrogen receptor-positive, HER2-negative, advanced breast cancer (PALOMA-1/TRIO-18): a randomised phase 2 study. Lancet Oncol. 2015;16(1):25–35. doi: 10.1016/S1470-2045(14)71159-3. [Finn RS, Crown JP, Lang I, et al. The cyclin-dependent kinase 4/6 inhibitor palbociclib in combination with letrozole versus letrozole alone as first-line treatment of oestrogen receptor-positive, HER2-negative, advanced breast cancer (PALOMA-1/TRIO-18): a randomised phase 2 study[J]. Lancet Oncol, 2015, 16(1): 25-35.] [DOI] [PubMed] [Google Scholar]
  • 8.Dickson MA. Molecular pathways: CDK4 inhibitors for cancer therapy. Clin Cancer Res. 2014;20(13):3379–83. doi: 10.1158/1078-0432.CCR-13-1551. [Dickson MA. Molecular pathways: CDK4 inhibitors for cancer therapy[J]. Clin Cancer Res, 2014, 20(13): 3379-83.] [DOI] [PubMed] [Google Scholar]
  • 9.Fernández-Aroca DM, Roche O, Sabater S, et al. P53 pathway is a major determinant in the radiosensitizing effect of Palbociclib: Implication in cancer therapy. Cancer Lett. 2019;451(5):23–33. doi: 10.1016/j.canlet.2019.02.049. [Fernández-Aroca DM, Roche O, Sabater S, et al. P53 pathway is a major determinant in the radiosensitizing effect of Palbociclib: Implication in cancer therapy[J]. Cancer Lett, 2019, 451(5): 23-33.] [DOI] [PubMed] [Google Scholar]
  • 10.Ge JY, Shu SK, Kwon M, et al. Acquired resistance to combined BET and CDK4/6 inhibition in triple-negative breast cancer. Nat Commun. 2020;11(1):2350. doi: 10.1038/s41467-020-16170-3. [Ge JY, Shu SK, Kwon M, et al. Acquired resistance to combined BET and CDK4/6 inhibition in triple-negative breast cancer[J]. Nat Commun, 2020, 11(1): 2350.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Jin H, Zhang Y, Ding Q, et al. Epithelial innate immunity mediates tubular cell senescence after kidney injury. JCI Insight. 2019;4(2):125490. doi: 10.1172/jci.insight.125490. [Jin H, Zhang Y, Ding Q, et al. Epithelial innate immunity mediates tubular cell senescence after kidney injury[J]. JCI Insight, 2019, 4 (2): 125490.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Canaud G, Brooks CR, Kishi S, et al. Cyclin G1 and TASCC regulate kidney epithelial cell G2-M arrest and fibrotic maladaptive repair. Sci Transl Med. 2019;11(476):eaav4754. doi: 10.1126/scitranslmed.aav4754. [Canaud G, Brooks CR, Kishi S, et al. Cyclin G1 and TASCC regulate kidney epithelial cell G2-M arrest and fibrotic maladaptive repair[J]. Sci Transl Med, 2019, 11(476): eaav4754.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Perazella MA. Onco-nephrology: renal toxicities of chemotherapeutic agents. Clin J Am Soc Nephrol. 2012;7(10):1713–21. doi: 10.2215/CJN.02780312. [Perazella MA. Onco-nephrology: renal toxicities of chemotherapeutic agents[J]. Clin J Am Soc Nephrol, 2012, 7(10): 1713-21.] [DOI] [PubMed] [Google Scholar]
  • 14.Cosmai L, Porta C, Gallieni M, et al. Onco-nephrology: a Decalogue. Nephrol Dial Transplant. 2016;31(4):515–9. doi: 10.1093/ndt/gfv320. [Cosmai L, Porta C, Gallieni M, et al. Onco-nephrology: a Decalogue[J]. Nephrol Dial Transplant, 2016, 31(4): 515-9.] [DOI] [PubMed] [Google Scholar]
  • 15.Capasso A, Benigni A, Capitanio U, et al. Summary of the International Conference on Onco-Nephrology: an emerging field in medicine. Kidney Int. 2019;96(3):555–67. doi: 10.1016/j.kint.2019.04.043. [Capasso A, Benigni A, Capitanio U, et al. Summary of the International Conference on Onco-Nephrology: an emerging field in medicine[J]. Kidney Int, 2019, 96(3): 555-67.] [DOI] [PubMed] [Google Scholar]
  • 16.Turner NC, Ro J, André F, et al. Palbociclib in hormone-receptorpositive advanced breast cancer. N Engl J Med. 2015;373(3):209–19. doi: 10.1056/NEJMoa1505270. [Turner NC, Ro J, André F, et al. Palbociclib in hormone-receptorpositive advanced breast cancer[J]. N Engl J Med, 2015, 373(3): 209-19.] [DOI] [PubMed] [Google Scholar]
  • 17.DeMichele A, Clark AS, Tan KS, et al. CDK4 /6 inhibitor palbociclib (PD0332991) in Rb + advanced breast cancer: phase Ⅱ activity, safety, and predictive biomarker assessment. Clin Cancer Res. 2015;21(5):995–1001. doi: 10.1158/1078-0432.CCR-14-2258. [DeMichele A, Clark AS, Tan KS, et al. CDK4 /6 inhibitor palbociclib (PD0332991) in Rb + advanced breast cancer: phase Ⅱ activity, safety, and predictive biomarker assessment[J]. Clin Cancer Res, 2015, 21(5): 995-1001.] [DOI] [PubMed] [Google Scholar]
  • 18.Zhang BY, Lam EW, Sun Y. Senescent cells: a new Achilles' heel to exploit for cancer medicine? Aging Cell. 2019;18(1):e12875. doi: 10.1111/acel.12875. [Zhang BY, Lam EW, Sun Y. Senescent cells: a new Achilles' heel to exploit for cancer medicine?[J]. Aging Cell, 2019, 18(1): e12875.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Ogrodnik M, Salmonowicz H, Jurk D, et al. Expansion and cellcycle arrest: common denominators of cellular senescence. Trends Biochem Sci. 2019;44(12):996–1008. doi: 10.1016/j.tibs.2019.06.011. [Ogrodnik M, Salmonowicz H, Jurk D, et al. Expansion and cellcycle arrest: common denominators of cellular senescence[J]. Trends Biochem Sci, 2019, 44(12): 996-1008.] [DOI] [PubMed] [Google Scholar]
  • 20.Klein ME, Kovatcheva M, Davis LE, et al. CDK4/6 inhibitors: the mechanism of action may not be as simple as once thought. Cancer Cell. 2018;34(1):9–20. doi: 10.1016/j.ccell.2018.03.023. [Klein ME, Kovatcheva M, Davis LE, et al. CDK4/6 inhibitors: the mechanism of action may not be as simple as once thought[J]. Cancer Cell, 2018, 34(1): 9-20.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Yuan LF, Alexander PB, Wang XF. Cellular senescence: from anticancer weapon to anti-aging target. Sci China Life Sci. 2020;63(3):332–42. doi: 10.1007/s11427-019-1629-6. [Yuan LF, Alexander PB, Wang XF. Cellular senescence: from anticancer weapon to anti-aging target[J]. Sci China Life Sci, 2020, 63 (3): 332-42.] [DOI] [PubMed] [Google Scholar]
  • 22.Kishino E, Ogata R, Saitoh W, et al. Anti-cell growth and anticancer stem cell activity of the CDK4/6 inhibitor palbociclib in breast cancer cells. Breast Cancer. 2020;27(3):415–25. doi: 10.1007/s12282-019-01035-5. [Kishino E, Ogata R, Saitoh W, et al. Anti-cell growth and anticancer stem cell activity of the CDK4/6 inhibitor palbociclib in breast cancer cells[J]. Breast Cancer, 2020, 27(3): 415-25.] [DOI] [PubMed] [Google Scholar]
  • 23.Chung L, Maestas DR Jr, Lebid A, et al. Interleukin 17 and senescent cells regulate the foreign body response to synthetic material implants in mice and humans. Sci Transl Med. 2020;12(539):eaax3799. doi: 10.1126/scitranslmed.aax3799. [Chung L, Maestas DR Jr, Lebid A, et al. Interleukin 17 and senescent cells regulate the foreign body response to synthetic material implants in mice and humans[J]. Sci Transl Med, 2020, 12 (539): eaax3799.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Liu JY, Souroullas GP, Diekman BO, et al. Cells exhibiting strong p16INK4a promoter activation in vivo display features of senescence. Proc Natl Acad Sci USA. 2019;116(7):2603–11. doi: 10.1073/pnas.1818313116. [Liu JY, Souroullas GP, Diekman BO, et al. Cells exhibiting strong p16INK4a promoter activation in vivo display features of senescence[J]. Proc Natl Acad Sci USA, 2019, 116(7): 2603-11.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25.Victorelli S, Lagnado A, Halim J, et al. Senescent human melanocytes drive skin ageing via paracrine telomere dysfunction. EMBO J. 2019;38(23):e101982. doi: 10.15252/embj.2019101982. [Victorelli S, Lagnado A, Halim J, et al. Senescent human melanocytes drive skin ageing via paracrine telomere dysfunction[J]. EMBO J, 2019, 38(23): e101982.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Chambers ES, Akbar AN. Can blocking inflammation enhance immunity during aging? J Allergy Clin Immunol. 2020;145(5):1323–31. doi: 10.1016/j.jaci.2020.03.016. [Chambers ES, Akbar AN. Can blocking inflammation enhance immunity during aging?[J]. J Allergy Clin Immunol, 2020, 145(5): 1323-31.] [DOI] [PubMed] [Google Scholar]
  • 27.Lehmann J, Baar MP, de Keizer PLJ. Senescent cells drive frailty through systemic signals. Trends Mol Med. 2018;24(11):917–8. doi: 10.1016/j.molmed.2018.09.003. [Lehmann J, Baar MP, de Keizer PLJ. Senescent cells drive frailty through systemic signals[J]. Trends Mol Med, 2018, 24(11): 917-8.] [DOI] [PubMed] [Google Scholar]
  • 28.Ritschka B, Storer M, Mas A, et al. The senescence-associated secretory phenotype induces cellular plasticity and tissue regeneration. Genes Dev. 2017;31(2):172–83. doi: 10.1101/gad.290635.116. [Ritschka B, Storer M, Mas A, et al. The senescence-associated secretory phenotype induces cellular plasticity and tissue regeneration[J]. Genes Dev, 2017, 31(2): 172-83.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29.Milanović V, Osimani A, Roncolini A, et al. Investigation of the dominant microbiota in ready-to-eat grasshoppers and mealworms and quantification of carbapenem resistance genes by QPCR. Front Microbiol. 2018;9:3036. doi: 10.3389/fmicb.2018.03036. [Milanović V, Osimani A, Roncolini A, et al. Investigation of the dominant microbiota in ready-to-eat grasshoppers and mealworms and quantification of carbapenem resistance genes by QPCR[J]. Front Microbiol, 2018, 9: 3036.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Barnes PJ, Baker J, Donnelly LE. Cellular senescence as a mechanism and target in chronic lung diseases. Am J Respir Crit Care Med. 2019;200(5):556–64. doi: 10.1164/rccm.201810-1975TR. [Barnes PJ, Baker J, Donnelly LE. Cellular senescence as a mechanism and target in chronic lung diseases[J]. Am J Respir Crit Care Med, 2019, 200(5): 556-64.] [DOI] [PubMed] [Google Scholar]
  • 31.Sturmlechner I, Durik M, Sieben CJ, et al. Cellular senescence in renal ageing and disease. Nat Rev Nephrol. 2017;13(2):77–89. doi: 10.1038/nrneph.2016.183. [Sturmlechner I, Durik M, Sieben CJ, et al. Cellular senescence in renal ageing and disease[J]. Nat Rev Nephrol, 2017, 13(2): 77-89.] [DOI] [PubMed] [Google Scholar]
  • 32.Qi RC, Yang C. Renal tubular epithelial cells: the neglected mediator of tubulointerstitial fibrosis after injury. Cell Death Dis. 2018;9(11):1126. doi: 10.1038/s41419-018-1157-x. [Qi RC, Yang C. Renal tubular epithelial cells: the neglected mediator of tubulointerstitial fibrosis after injury[J]. Cell Death Dis, 2018, 9 (11): 1126.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Schafer MJ, White TA, Iijima K, et al. Cellular senescence mediates fibrotic pulmonary disease. Nat Commun. 2017;8:14532. doi: 10.1038/ncomms14532. [Schafer MJ, White TA, Iijima K, et al. Cellular senescence mediates fibrotic pulmonary disease[J]. Nat Commun, 2017, 8: 14532.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34.Wiley CD, Velarde MC, Lecot P, et al. Mitochondrial dysfunction induces senescence with a distinct secretory phenotype. Cell Metab. 2016;23(2):303–14. doi: 10.1016/j.cmet.2015.11.011. [Wiley CD, Velarde MC, Lecot P, et al. Mitochondrial dysfunction induces senescence with a distinct secretory phenotype[J]. Cell Metab, 2016, 23(2): 303-14.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Birch J, Passos JF. Targeting the SASP to combat ageing: Mitochondria as possible intracellular allies? Bioessays. 2017;39:1600235. doi: 10.1002/bies.201600235. [Birch J, Passos JF. Targeting the SASP to combat ageing: Mitochondria as possible intracellular allies?[J]. Bioessays, 2017, 39: 1600235.] [DOI] [PubMed] [Google Scholar]
  • 36.Ferenbach DA, Bonventre JV. Mechanisms of maladaptive repair after AKI leading to accelerated kidney ageing and CKD. Nat Rev Nephrol. 2015;11(5):264–76. doi: 10.1038/nrneph.2015.3. [Ferenbach DA, Bonventre JV. Mechanisms of maladaptive repair after AKI leading to accelerated kidney ageing and CKD[J]. Nat Rev Nephrol, 2015, 11(5): 264-76.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37.Xiao LX, Zhou D, Tan RJ, et al. Sustained activation of wnt/β-catenin signaling drives AKI to CKD progression. J Am Soc Nephrol. 2016;27(6):1727–40. doi: 10.1681/ASN.2015040449. [Xiao LX, Zhou D, Tan RJ, et al. Sustained activation of wnt/β-catenin signaling drives AKI to CKD progression[J]. J Am Soc Nephrol, 2016, 27(6): 1727-40.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 38.Andrade L, Rodrigues CE, Gomes SA, et al. Acute kidney injury as a condition of renal senescence. Cell Transplant. 2018;27(5):739–53. doi: 10.1177/0963689717743512. [Andrade L, Rodrigues CE, Gomes SA, et al. Acute kidney injury as a condition of renal senescence[J]. Cell Transplant, 2018, 27(5): 739-53.] [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Li CZ, Xie N, Li Y, et al. N-acetylcysteine ameliorates cisplatininduced renal senescence and renal interstitial fibrosis through sirtuin1 activation and p53 deacetylation. Free Radic Biol Med. 2019;130:512–27. doi: 10.1016/j.freeradbiomed.2018.11.006. [Li CZ, Xie N, Li Y, et al. N-acetylcysteine ameliorates cisplatininduced renal senescence and renal interstitial fibrosis through sirtuin1 activation and p53 deacetylation[J]. Free Radic Biol Med, 2019, 130: 512-27.] [DOI] [PubMed] [Google Scholar]

Articles from Journal of Southern Medical University are provided here courtesy of Editorial Department of Journal of Southern Medical University

RESOURCES