應(yīng)用紀(jì)要：上半年Incucyte國內(nèi)成果大盤點(diǎn)

陳老濕最近又迷上一款革命性的細(xì)胞成像設(shè)備-- Incucyte^® 長時(shí)間實(shí)時(shí)活細(xì)胞成像系統(tǒng)，國內(nèi)這款儀器用得怎么樣呢？讓陳老濕細(xì)細(xì)道來！去除預(yù)印版和綜述，2022年上半年使用Incucyte^® 發(fā)表的文章有大幾十篇（文獻(xiàn)列表見文末），僅第一作者就覆蓋了至少40個(gè)單位。
 
以下是發(fā)表至少2篇以上文章的單位列表：

• 澳門大學(xué)中醫(yī)研究院（4篇）
• 復(fù)旦大學(xué)上海癌癥中心（3篇）
• 中科院動(dòng)物研究所（3篇）
• 濱州醫(yī)學(xué)院（2篇）
• 鄭州大學(xué)基礎(chǔ)醫(yī)學(xué)院（2篇）
• 中山大學(xué)附屬第六醫(yī)院（2篇）

 
可見，很多單位可以利用Incucyte^® 實(shí)時(shí)、高效地獲得細(xì)胞信息和可信的研究成果，陳老濕去一些單位拜訪的時(shí)候，發(fā)現(xiàn)這臺(tái)儀器都是搶著使用的，原因結(jié)尾會(huì)總結(jié)，首先一點(diǎn)就是Incucyte^® 應(yīng)用非常廣泛，從這些文章上看，最多的應(yīng)用是細(xì)胞增殖 & 細(xì)胞活性實(shí)驗(yàn)，然后是劃痕遷移實(shí)驗(yàn)，當(dāng)然還有3D拍攝、神經(jīng)細(xì)胞分析、吞噬等亮點(diǎn)應(yīng)用。
 
陳老濕就選幾篇和大家嘮一下：

 
 
Incucyte^® 多點(diǎn)開花應(yīng)用
山東齊魯醫(yī)院
癌癥治療新靶點(diǎn)^【1】

ETS 轉(zhuǎn)錄因子 GABPA 長期以來一直被認(rèn)為是一種致癌因子，最近有人提出由于其對端粒酶活化的關(guān)鍵作用，可以作為癌癥治療的靶點(diǎn)。但 GABPA對細(xì)胞腎細(xì)胞癌（ccRCC）形成的機(jī)理尚不清楚, 利用 siRNA 和表達(dá)載體對 GABPA 等因子的表達(dá)進(jìn)行調(diào)控發(fā)現(xiàn)，GABPA 耗竭雖然抑制端粒酶表達(dá)，但顯著增強(qiáng)了增殖、侵襲和ccRCC 細(xì)胞的干性，而 GABPA 過表達(dá)表現(xiàn)出相反的作用，強(qiáng)烈抑制體內(nèi)轉(zhuǎn)移和致癌。TGFBR2 被鑒定為 GABPA 靶基因，GABPA 通過它控制 TGFβ信號(hào)控制ccRCC 表型。Incucyte^® 被用于各種細(xì)胞表征實(shí)驗(yàn)。


細(xì)胞劃痕遷移實(shí)驗(yàn)：證明GABPA過表達(dá)會(huì)抑制ccRCC細(xì)胞系（A498，786-O）的遷移
  二維細(xì)胞增殖實(shí)驗(yàn)（左）和三維腫瘤球?qū)嶒?yàn)（D,E）表明，沉默GABPA表達(dá)加強(qiáng)ccRCC細(xì)胞系（A498，786-O）的增殖（D圖），GABPA載體增加表達(dá)則減弱增殖（E圖）。
  
劃痕實(shí)驗(yàn)，2D和3D拍攝，Incucyte^®都在行！
 
 
Incucyte^®的高通量
中科院動(dòng)物研究所
干細(xì)胞防衰老劑的小分子篩選^【2】

使用 Incucyte^® S3活細(xì)胞成像系統(tǒng)，通過監(jiān)測上千種 FDA 批準(zhǔn)的藥物對Werner綜合征間充質(zhì)干細(xì)胞 （WS hMSCs） 的增殖活性影響，建立了一種高通量藥物篩選方法。1,622 種 FDA 批準(zhǔn)的藥物在一定濃度下持續(xù)作用于WS hMSCs 6天。通過初步篩選，確定了前 20 名候選藥物。這些候選藥物包括卡博替尼、比卡魯胺、沒食子酸 (GA)等。GA 是一種天然酚類化合物，廣泛存在于茶、水果和酒中。通過功能性實(shí)驗(yàn)，發(fā)現(xiàn)GA作為干細(xì)胞防衰老劑的活性比二甲雙胍和槲皮素還要好。
 

用Incucyte^® 計(jì)算藥物處理6天后的細(xì)胞匯合度的與對照的比值，值越大說明藥物增強(qiáng)細(xì)胞增殖的能力越強(qiáng)。
 
Incucyte^®一次性可以放6個(gè)孔板，其通量可以滿足上千個(gè)化合物的篩選！
 
 
Incucyte^®吞噬作用
南京中醫(yī)藥大學(xué)
天然產(chǎn)物AD藥物^【3】

臨床證據(jù)表明，絕經(jīng)后女性患阿爾茨海默病 (AD) 的可能性幾乎是同齡男性的兩倍，雌激素與AD的發(fā)生密切相關(guān)。雌激素受體ERβ 主要分布在海馬體和心血管系統(tǒng)中，ERβ 選擇性激動(dòng)劑是一種有價(jià)值的抗神經(jīng)退行性疾病的藥物。在這項(xiàng)研究中，確定了一種天然產(chǎn)物廣藿香醇 (PTA) 作為選擇性 ERβ激動(dòng)劑改善雌性 APP/PS1 小鼠的認(rèn)知缺陷，并探索其潛在機(jī)制。小鼠體內(nèi)實(shí)驗(yàn)發(fā)現(xiàn)PTA通過促進(jìn)小膠質(zhì)細(xì)胞的吞噬作用，減少海馬中的淀粉樣斑塊沉積；在體外也觀察到 PTA 的治療效果：PTA (5, 10, 20 μM) 劑量依賴性增加原代小膠質(zhì)細(xì)胞和 BV2 細(xì)胞中 o-FAM-Aβ42 的吞噬作用，本研究探討了選擇性 ERβ 激動(dòng)劑在改善 AD 并突出 PTA 作為抗疾病藥物先導(dǎo)化合物的潛力。
  PTA可以增強(qiáng)BV2細(xì)胞對FAM標(biāo)記的Aβ42的吞噬（綠色熒光增多，上面兩圖以及圖d），但是不影響Aβ42的降解（圖e）。
 
Incucyte^®有各種配套試劑，可以滿足吞噬，內(nèi)吞，代謝等實(shí)驗(yàn)。
 
 

• 培養(yǎng)箱內(nèi)可長達(dá)數(shù)周的連續(xù)觀察，最短幾分鐘間隔拍攝，減少人力，防止過多操作對細(xì)胞的傷害；
• 6個(gè)板位，分別獨(dú)立設(shè)置檢測程序，可以兼容各種孔板和培養(yǎng)皿，通量高；
• 高效簡便的模塊化軟件設(shè)置和數(shù)據(jù)分析，輸出圖片、視頻、生長曲線等多指標(biāo)多參數(shù)；
• 大于100種優(yōu)化過的熒光試劑和耗材及詳盡的Protocol.

   
 
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-2022年上半年使用Incucyte^® 發(fā)表的文章 -
 

1. Myricetin inhibits interferon-γ-induced PD-L1 and IDO1 expression in lung cancer cells.Biochemical Pharmacology 197 (2022) 114940
2. 17β-estradiol rescues the damage of thiazolidinedione on chicken Sertoli cell proliferation via adiponectin.Ecotoxicology and Environmental Safety 233 (2022) 113308
3. 3,30-Diindolylmethane Enhances Fluorouracil Sensitivity via Inhibition of Pyrimidine Metabolism in Colorectal Cancer.Metabolites 2022,12, 410.
4. An optimized method for obtaining clinical-grade specific cell subpopulations from human umbilical cord-derived mesenchymal stem cells.Cell Prolif. 2022;e13300.
5. ATF3 -activated accelerating effect of LINC00941/lncIAPF on fibroblast-to-myofibroblast differentiation by blocking autophagy depending on ELAVL1/HuR in pulmonary fibrosis.AUTOPHAGY https://doi.org/10.1080/15548627.2022.2046448
6. Cholecystectomy promotes colon carcinogenesis by activating the Wnt signaling pathway by increasing the deoxycholic acid level Center.Cell Communication and Signaling 2022, 20(1):71
7. Correction to: Downregulation of fibronectin 1 attenuates ATRA‑induced inhibition of cell migration and invasion in neuroblastoma cells.Molecular and Cellular Biochemistry (2022) 477:1323–1326
8. Cross-species metabolomic analysis identifies uridine as a potent regeneration promoting factor.Cell Discovery ( 2022) 8:6
9. Cytochrome P450 26A1 regulates the clusters and killing activity of NK cells during the peri-implantation period.J Cell Mol Med. 2022;26:2438–2450.
10. DGKZ promotes TGFβ signaling pathway and metastasis in triple-negative breast cancer by suppressing lipid raft-dependent endocytosis of TGFβR2.Cell Death and Disease (2022) 13:105
11. Dysregulated lipid metabolism blunts the sensitivity of cancer cells to EZH2 inhibitor.eBioMedicine 2022;77:103872
12. Emodin targeting the colonic metabolism via PPARγ alleviates UC by inhibiting facultative anaerobe.Phytomedicine 104 (2022) 154106 Available
13. Exogenous iron impairs the anti-cancer effect of ascorbic acid both in vitro and in vivo.Journal of Advanced Research
14. Fluoride exposure induces lysosomal dysfunction unveiled by an integrated transcriptomic and metabolomic study in bone marrow mesenchymal stem cells.Ecotoxicology and Environmental Safety 239 (2022) 113672
15. GABPA‑activated TGFBR2 transcription inhibits aggressiveness but is epigenetically erased by oncometabolites in renal cell carcinoma.J Exp Clin Cancer Res (2022) 41:173
16. Genome-scale CRISPR–Cas9 screen reveals novel regulators of B7-H3 in tumor cells.J Immunother Cancer 2022;10:e004875.
17. Glycopolymer Engineering of the Cell Surface Changes the Single Cell Migratory Direction and Inhibits the Collective Migration of Cancer Cells.ACS Appl. Mater. Interfaces 2022, 14, 4921−4930
18. hnRNPL-activated circANKRD42 back-splicing and circANKRD42-mediated crosstalk of mechanical stiffness and biochemical signal in lung fibrosis.Molecular Therapy Vol. 30 No 6
19. HTR1A Inhibits the Progression of Triple-Negative Breast Cancer via TGF-𝜷 Canonical and Noncanonical Pathways.Adv. Sci. 2022, 9, 2105672
20. Human lysyl‑tRNA synthetase evolves a dynamic structure that can be stabilized by forming complex.Cellular and Molecular Life Sciences (2022) 79:128
21. Industrially synthesized biosafe vaterite hollow CaCO3 for controllable delivery of anticancer drugs.Materials Today Chemistry 24 (2022) 100917
22. Knockdown of Peroxiredoxin V increased the cytotoxicity of non-thermal plasma-treated culture medium to A549 cells.AGING 2022
23. Large-scale chemical screen identifies Gallic acid as a geroprotector for human stem cells.Protein Cell 2022, 13(7):532–539
24. Ligand-Directed Caging Enables the Control of Endogenous DNA Alkyltransferase Activity with Light inside Live Cells.Angew. Chem. Int. Ed. 2022
25. Limiting dilution assay to quantify the self-renewal potential of cancer stem cells in hepatocellular carcinoma.Methods in Cell Biology, ISSN 0091-679X, https://doi.org/10.1016/bs.mcb.2022.04.010
26. Long non‑coding RNA LSAMP‑1 is down‑regulated in non‑small cell lung cancer and predicts a poor prognosis.Cancer Cell International (2022) 22:181
27. Low doses of niclosamide and quinacrine combination yields synergistic effect in melanoma via activating autophagy-mediated p53-dependent apoptosis.Translational Oncology 21 (2022) 101425
28. Low-dose chloroquine treatment extends the lifespan of aged rats.Protein Cell 2022, 13(6):454–461
29. Mutations and clinical significance of calcium voltage-gated channel subunit alpha 1E (CACNA1E) in non-small cell lung cancer.https://doi.org/10.1016/j.annonc.2022.02.208
30. Network pharmacology and in vitro studies reveal the pharmacological effects and molecular mechanisms of Shenzhi Jiannao prescription against vascular dementia.BMC Complementary Medicine and Therapies (2022) 22:33
31. Nonmetal Graphdiyne Nanozyme-Based Ferroptosis−Apoptosis Strategy for Colon Cancer Therapy.ACS Appl. Mater. Interfaces 2022, 14, 27720−27732
32. Novel prognostic marker LINC00205 promotes tumorigenesis and metastasis by competitively suppressing miRNA-26a in gastric cancer.Cell Death Discovery (2022) 8:5
33. Patchouli alcohol as a selective estrogen receptor β agonist ameliorates AD-like pathology of APP/PS1 model mice.Acta Pharmacologica Sinica (2022) 0:1–16
34. Promoting role of pentraxin-3 in esophageal squamous cell carcinoma.Molecular Therapy
35. Psoralidin, a natural compound from Psoralea corylifolia, induces oxidative damage mediated apoptosis in colon cancer cells.J Biochem Mol Toxicol. 2022;e23051
36. Receptor activity‐modifying protein 1 regulates mouse skin fibroblast proliferation via the Gαi3‑PKA‑CREB‑YAP axis.Cell Communication and Signaling (2022) 20:52
37. Redirecting anti-Vaccinia virus T cell immunity for cancer treatment by AAV-mediated delivery of the VV B8R gene.Molecular Therapy
38. Research on selective uptake of photosensitizer C3N4@RP by different cancer cells.Mater. Res. Express 9 (2022) 065402
39. RNF4 silencing induces cell growth arrest and DNA damage by promoting nuclear targeting of p62 in hepatocellular carcinoma.Oncogene (2022) 41:2275–2286
40. S100A9-CXCL12 activation in BRCA1-mutant breast cancer promotes an immunosuppressive microenvironment associated with resistance to immunotherapy.NATURE COMMUNICATIONS | (2022) 13:1481
41. Schlafen 5 suppresses human immunodeficiency virus type 1 transcription by commandeering cellular epigenetic machinery.Nucleic Acids Research, 2022
42. Shuxuetong injection and its peptides enhance angiogenesis after hindlimb ischemia by activating the MYPT1/LIMK1/Cofilin pathway.Journal of Ethnopharmacology 292 (2022) 115166
43. SIK2 promotes ovarian cancer cell motility and metastasis by phosphorylating MYLK.Molecular Oncology 16 (2022) 2558–2574
44. SLC1A1-mediated cellular and mitochondrial influx of R-2-hydroxyglutarate in vascular endothelial cells promotes tumor angiogenesis in IDH1-mutant solid tumors.Cell Research (2022) 32:638–658
45. SLC35E2 promoter mutation as a prognostic marker of esophageal squamous cell carcinoma.Life Sciences 296 (2022) 120447
46. Suppression of toxicity of the mutant huntingtin protein by its interacting compound, desonide.PNAS 119 (10) e2114303119
47. Systematic Survey of the Regulatory Networks of the Long Noncoding RNA BANCR in Cervical Cancer Cells.J. Proteome Res. 2022, 21, 1137−1152
48. The early‑stage triple‑negative breast cancer landscape derives a novel prognostic signature and therapeutic target.Breast Cancer Research and Treatment (2022) 193:319–330
49. The SF3B1R625H mutation promotes prolactinoma tumor progression through aberrant splicing of DLG1.J Exp Clin Cancer Res (2022) 41:26
50. The two‑faced role of ATF2 on cisplatin response in gastric cancer depends on p53 context.Cell & Bioscience (2022) 12:77
51. Toosendanin, a late‑stage autophagy inhibitor, sensitizes triple‑negative breast cancer to irinotecan chemotherapy.Chinese Medicine (2022) 17:55
52. TSG-6 promotes Cancer Cell aggressiveness in a CD44-Dependent Manner and Reprograms Normal Fibroblasts to create a Pro-metastatic Microenvironment in Colorectal Cancer.Int. J. Biol. Sci. 2022, Vol. 18
53. Upregulation of ADAMDEC1 correlates with tumor progression and predicts poor prognosis in non-small cell lung cancer (NSCLC) via the PI3K/AKT pathway.Thorac Cancer. 2022;13:1027–1039

-參考文獻(xiàn)-
【1】GABPA‑activated TGFBR2 transcription inhibits aggressiveness but is epigenetically erased by oncometabolites in renal cell carcinoma.J Exp Clin Cancer Res (2022) 41:173
【2】Large-scale chemical screen identifies Gallic acid as a geroprotector for human stem cells.Protein Cell 2022, 13(7):532–539
【3】Patchouli alcohol as a selective estrogen receptor β agonist ameliorates AD-like pathology of APP/PS1 model mice.Acta Pharmacologica Sinica (2022) 0:1–16