Citation: PANG Shu-Yun, XU Shu, XU Zhao-Luo, CHU Ya-Nan, MA Xue-Ping, WU Hai-Ping, ZOU Bing-Jie, ZHOU Guo-Hua. Quality Evaluation of Capture and Single-cell Isolation of Circulating Tumor Cell Clusters for Single-cell Ribonucleic Acid Analysis[J]. Chinese Journal of Analytical Chemistry, ;2023, 51(4): 513-520. doi: 10.19756/j.issn.0253-3820.221526 shu

Quality Evaluation of Capture and Single-cell Isolation of Circulating Tumor Cell Clusters for Single-cell Ribonucleic Acid Analysis

PANG Shu-Yun ¹ ,
XU Shu ⁵ ,
XU Zhao-Luo ¹ ,
CHU Ya-Nan ¹ ,
MA Xue-Ping ¹ ,
WU Hai-Ping ^1,3 ,
ZOU Bing-Jie ^1,3,4,, ,
ZHOU Guo-Hua ^1,2,3,,

1.
Department of Clinical Pharmacy, Jinling Hospital, Medical School of Nanjing University, Nanjing 210002, China;
2.
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China;
3.
School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China;
4.
School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China;
5.
School of Basic Medical Science and Clinical Pharmacy, China Pharmaceutical University, Nanjing 210009, China

Corresponding author: ZOU Bing-Jie, ZHOU Guo-Hua,
Received Date: 25 October 2022
Revised Date: 28 December 2022

Fund Project: Supported by the National Natural Science Foundation of China (No. 61871403).

Single-cell ribonucleic acid (RNA) analysis is necessary to investigate the biological effects of circulating tumor cell clusters (CTC clusters). To achieve the most efficient results in single-cell analysis of CTC clusters, the integrity of the CTC cluster and the high quality of the intracellular RNA should be maintained after the capture and single-cell isolation. So, three house gene was quantitated in single cells for RNA quality control by single-cell RNA sequencing method, and the CTC cluster's lateral diameter for evaluating the CTC cluster's integrity were used to evaluate the quality of CTC cluster capture and single-cell isolation process. Trypsin-EDTA and non-enzymatic cell dissociation solution (NECDS) were used to isolate single cells in CTC clusters, followed by the investigation of intracellular RNA quality. The CTC cluster recovery of negative enrichment, positive enrichment and membrane filtration, and single-cell RNA integrity after enrichment were investigated. The results indicated that the NECDS isolated single cells from CTC clusters rapidly without affecting the intracellular RNA. Besides, it was found that rotational incubation and treatments of red blood cell lysate disrupted the integrity of CTC clusters, therefore, these operations were not the best way for single-cell RNA analysis of CTC clusters. The positive enrichment with simplified process achieved a high recovery without affecting the quality of intracellular RNA. Single-cell RNA analysis of CTC clusters required gentle capture and single-cell isolation process. The present study here provided an efficient sample preparation strategy for single-cell analysis of CTC clusters.
- Circulating tumor cell clusters,
- Single-cell ribonucleic acid analysis,
- Single-cell isolation,
- Cluster capture
1. [1]
  ACETO N, BARDIA A, MIYAMOTO D T, DONALDSON M C, WITTNER B S, SPENCER J A, YU M, PELY A, ENGSTROM A, ZHU H, BRANNIGAN B W, KAPUR R, STOTT S L, SHIODA T, RAMASWAMY S, TING D T, LIN C P, TONER M, HABER D A, MAHESWARAN S. Cell, 2014, 158(5):1110-1122.
2. [2]
  CHEUNG K J, EWALD A J. Science, 2016, 352(6282):167-169.
3. [3]
  GKOUNTELA S, CASTRO-GINER F, SZCZERBA B M, VETTER M, LANDIN J, SCHERRER R, KROL I, SCHEIDMANN M C, BEISEL C, STIRNIMANN C U, KURZEDER C, HEINZELMANN-SCHWARZ V, ROCHLITZ C, WEBER W P, ACETO N. Cell, 2019, 176(1-2):98-112.
4. [4]
  COSTA C, MUINELO-ROMAY L, CEBEY-LÓPEZ V, PEREIRA-VEIGA T, MARTÍNEZ-PENA I, ABREU M, ABALO A, LAGO-LESTÓN R M, ABUÍN C, PALACIOS P, CUEVA J, PIÑEIRO R, LÓPEZ-LÓPEZ R. Cancers, 2020, 12(5):1111.
5. [5]
  SZCZERBA B M, CASTRO-GINER F, VETTER M, KROL I, GKOUNTELA S, LANDIN J, SCHEIDMANN M C, DONATO C, SCHERRER R, SINGER J, BEISEL C, KURZEDER C, HEINZELMANN-SCHWARZ V, ROCHLITZ C, WEBER W P, BEERENWINKEL N, ACETO N. Nature, 2019, 566(7745):553-557.
6. [6]
  LIU X, TAFTAF R, KAWAGUCHI M, CHANG Y F, CHEN W, ENTENBERG D, ZHANG Y, GERRATANA L, HUANG S, PATEL D B, TSUI E, ADORNO-CRUZ V, CHIRIELEISON S M, CAO Y, HARNEY A S, PATEL S, PATSIALOU A, SHEN Y, AVRIL S, GILMORE H L, LATHIA J D, ABBOTT D W, CRISTOFANILLI M, CONDEELIS J S, LIU H. Cancer Discovery, 2019, 9(1):96-113.
7. [7]
  GRILLET F, BAYET E, VILLERONCE O, ZAPPIA L, LAGERQVIST E L, LUNKE S, CHARAFE-JAUFFRET E, PHAM K, MOLCK C, ROLLAND N, BOURGAUX J F, PRUDHOMME M, PHILIPPE C, BRAVO S, BOYER J C, CANTERELTHOUENNON L, TAYLOR G R, HSU A, PASCUSSI J M, HOLLANDE F, PANNEQUIN J. Gut, 2017, 66(10):1802-1810.
8. [8]
  MASSAGUé J, OBENAUF A C. Nature, 2016, 529(7586):298-306.
9. [9]
  JANSSON S, BENDAHL P O, LARSSON A M, AALTONEN K E, RYDÉN L. BMC Cancer, 2016, 16(1):433.
10. [10]
  ACETO N, TONER M, MAHESWARAN S, HABER D A. Trends Cancer, 2015, 1(1):44-52.
11. [11]
  TAFTAF R, LIU X, SINGH S, JIA Y, DASHZEVEG N K, HOFFMANN A D, EL-SHENNAWY L, RAMOS E K, ADORNO-CRUZ V, SCHUSTER E J, SCHOLTEN D, PATEL D, ZHANG Y, DAVIS A A, REDUZZI C, CAO Y, D'AMICO P, SHEN Y, CRISTOFANILLI M, MULLER W A, VARADAN V, LIU H. Nat. Commun., 2021, 12(1):4867.
12. [12]
  HONG M, TAO S, ZHANG L, DIAO L T, HUANG X, HUANG S, XIE S J, XIAO Z D, ZHANG H. J. Hematol. Oncol., 2020, 13(1):166.
13. [13]
  KOLODZIEJCZYK A A, KIM J K, SVENSSON V, MARIONI J C, TEICHMANN S A. Mol. Cell, 2015, 58(4):610-620.
14. [14]
  KIM S C, CLARK I C, SHAHI P, ABATE A R. Anal. Chem., 2018, 90(2):1273-1279.
15. [15]
  ZIEGENHAIN C, VIETH B, PAREKH S, REINIUS B, GUILLAUMET-ADKINS A, SMETS M, LEONHARDT H, HEYN H, HELLMANN I, ENARD W. Mol. Cell, 2017, 65(4):631-643.
16. [16]
  ZHANG Z, XIAO Y, ZHAO J, CHEN M, XU Y, ZHONG W, XING J, WANG M. Respirology, 2016, 21(3):519-525.
17. [17]
  WEN C Y, WU L L, ZHANG Z L, LIU Y L, WEI S Z, HU J, TANG M, SUN E Z, GONG Y P, YU J, PANG D W. ACS Nano, 2014, 8(1):941-949.
18. [18]
  TANG Y, SHI J, LI S, WANG L, CAYRE Y E, CHEN Y. Sci. Rep., 2014, 4(1):6052.
19. [19]
  MATSUNAGA H, GOTO M, ARIKAWA K, SHIRAI M, TSUNODA H, HUANG H, KAMBARA H. Anal. Biochem., 2015, 471:9-16.
20. [20]
  HUANG H, GOTO M, TSUNODA H, SUN L, TANIGUCHI K, MATSUNAGA H, KAMBARA H. Nucleic Acids Res., 2014, 42(2):e12.
21. [21]
  VRTAČNIK P, KOS Š, BUSTIN S A, MARC J, OSTANEK B. Anal. Biochem., 2014, 463:38-44.
22. [22]
  MU Z, WANG C, YE Z, AUSTIN L, CIVAN J, HYSLOP T, PALAZZO J P, JASLOW R, LI B, MYERS R E, JIANG J, XING J, YANG H, CRISTOFANILLI M. Breast Cancer Res. Treat., 2015, 154(3):563-571.
23. [23]
  FISCHER K R, DURRANS A, LEE S, SHENG J, LI F, WONG S T C, CHOI H, EL RAYES T, RYU S, TROEGER J, SCHWABE R F, VAHDAT L T, ALTORKI N K, MITTAL V, GAO D. Nature, 2015, 527(7579):472-476.
24. [24]
  ZHENG X, CARSTENS J L, KIM J, SCHEIBLE M, KAYE J, SUGIMOTO H, WU C C, LEBLEU V S, KALLURI R. Nature, 2015, 527(7579):525-530.
1. [1]
  Jiahao Zeng , Hui Chao . 诱导程序性细胞死亡的金属抗肿瘤药物研究. University Chemistry, 2025, 40(6): 145-159. doi: 10.12461/PKU.DXHX202406019
2. [2]
  Zhaoxin LI , Ruibo WEI , Min ZHANG , Zefeng WANG , Jing ZHENG , Jianbo LIU . Advancements in the construction of inorganic protocells and their cell mimic and bio-catalytical applications. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2286-2302. doi: 10.11862/CJIC.20240235
3. [3]
  Shuyu Liu , Xiaomin Sun , Bohan Song , Gaofeng Zeng , Bingbing Du , Chongshen Guo , Cong Wang , Lei Wang . Design and Fabrication of Phospholipid-Vesicle-based Artificial Cells towards Biomedical Applications. University Chemistry, 2024, 39(11): 182-188. doi: 10.12461/PKU.DXHX202404113
4. [4]
  Tiejin Chen , Xiaokuang Xue , Jian Li , Minhui Cui , Yongliang Hao , Mianqi Xue , Haihua Xiao , Jiechao Ge , Pengfei Wang . Membrane-anchoring nanoengineered carbon dots as a pyroptosis amplifier for robust tumor photodynamic-immunotherapy. Acta Physico-Chimica Sinica, 2025, 41(10): 100113-0. doi: 10.1016/j.actphy.2025.100113
5. [5]
  Siyi ZHONG , Xiaowen LIN , Jiaxin LIU , Ruyi WANG , Tao LIANG , Zhengfeng DENG , Ao ZHONG , Cuiping HAN . Targeting imaging and detection of ovarian cancer cells based on fluorescent magnetic carbon dots. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1483-1490. doi: 10.11862/CJIC.20240093
6. [6]
  Shipeng WANG , Shangyu XIE , Luxian LIANG , Xuehong WANG , Jie WEI , Deqiang WANG . Piezoelectric effect of Mn, Bi co-doped sodium niobate for promoting cell proliferation and bacteriostasis. Chinese Journal of Inorganic Chemistry, 2024, 40(10): 1919-1931. doi: 10.11862/CJIC.20240094
7. [7]
  Peng GENG , Guangcan XIANG , Wen ZHANG , Haichuang LAN , Shuzhang XIAO . Hollow copper sulfide loaded protoporphyrin for photothermal-sonodynamic therapy of cancer cells. Chinese Journal of Inorganic Chemistry, 2024, 40(10): 1903-1910. doi: 10.11862/CJIC.20240155
8. [8]
  Jie WEI , Qing ZHOU , Dandan DING , Xiang JING , Fei LI . Photothermal toxicity of Prussian blue nanoparticles to cervical cancer cells. Chinese Journal of Inorganic Chemistry, 2025, 41(11): 2345-2357. doi: 10.11862/CJIC.20240435
9. [9]
  Di WU , Ruimeng SHI , Zhaoyang WANG , Yuehua SHI , Fan YANG , Leyong ZENG . Construction of pH/photothermal dual-responsive delivery nanosystem for combination therapy of drug-resistant bladder cancer cell. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1679-1688. doi: 10.11862/CJIC.20240135
10. [10]
  Pengli GUAN , Renhu BAI , Xiuling SUN , Bin LIU . Trianiline-derived aggregation-induced emission luminogen probe for lipase detection and cell imaging. Chinese Journal of Inorganic Chemistry, 2025, 41(9): 1817-1826. doi: 10.11862/CJIC.20250058
11. [11]
  Jie ZHAO , Huili ZHANG , Xiaoqing LU , Zhaojie WANG . Theoretical calculations of CO₂ capture and separation by functional groups modified 2D covalent organic framework. Chinese Journal of Inorganic Chemistry, 2025, 41(2): 275-283. doi: 10.11862/CJIC.20240213
12. [12]
  Xinyu ZENG , Guhua TANG , Jianming OUYANG . Inhibitory effect of Desmodium styracifolium polysaccharides with different content of carboxyl groups on the growth, aggregation and cell adhesion of calcium oxalate crystals. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1563-1576. doi: 10.11862/CJIC.20230374
13. [13]
  Yanxi LIU , Mengjia XU , Haonan CHEN , Quan LIU , Yuming ZHANG . A fluorescent-colorimetric probe for peroxynitrite-anion-imaging in living cells. Chinese Journal of Inorganic Chemistry, 2025, 41(6): 1112-1122. doi: 10.11862/CJIC.20240423
14. [14]
  Lin LI , Jiaxue LI , Meixia YANG , Jiayu DING , Jiaqi JING , Ruiping ZHANG . Preparation of mitoxantrone self-assembled carrier-free nanodrugs regulated by sodium acetate for apoptosis induction of human breast carcinoma cells. Chinese Journal of Inorganic Chemistry, 2025, 41(12): 2536-2548. doi: 10.11862/CJIC.20250138
15. [15]
  Huan LI , Shengyan WANG , Long Zhang , Yue CAO , Xiaohan YANG , Ziliang WANG , Wenjuan ZHU , Wenlei ZHU , Yang ZHOU . Growth mechanisms and application potentials of magic-size clusters of groups Ⅱ-Ⅵ semiconductors. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1425-1441. doi: 10.11862/CJIC.20240088
16. [16]
  Xinyu Zhu , Meili Pang . Application of Functional Group Addition Strategy in Organic Synthesis. University Chemistry, 2024, 39(3): 218-230. doi: 10.3866/PKU.DXHX202308106
17. [17]
  Wen Jiang , Jieli Lin , Zhongshu Li . 低配位含磷官能团的研究进展. University Chemistry, 2025, 40(8): 138-151. doi: 10.12461/PKU.DXHX202409144
18. [18]
  Yuanyuan Ping , Wangqing Kong . 光催化碳氢键官能团化合成1-苯基-1,2-乙二醇. University Chemistry, 2025, 40(6): 238-247. doi: 10.12461/PKU.DXHX202408092
19. [19]
  Shengwen XU , Longlong YANG , Houji CAO , Deshuang TU , Xing WEI , Changsheng LU , Hong YAN . Research progress on light-induced functionalization of polyhedral carborane clusters. Chinese Journal of Inorganic Chemistry, 2025, 41(11): 2187-2200. doi: 10.11862/CJIC.20250192
20. [20]
  Qi Zhang , Ziyu Liu , Hongxia Tan , Jun Tong , Dazhen Xu . Research Progress on Direct Synthesis of β-Hydroxy Sulfones via Difunctionalization of Olefins. University Chemistry, 2025, 40(11): 199-209. doi: 10.12461/PKU.DXHX202412064

Get Citation

PDF

XML

Metrics

PDF Downloads(7)
Abstract views(1853)
HTML views(74)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142