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Self-targeted knockdown of CD44 improves cisplatin sensitivity of chemoresistant non-small cell lung cancer cells

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Abstract

Background

Chemoresistance remains a major challenge for effective chemotherapy of non-small-cell lung carcinoma (NSCLC). CD44 expression is related to the susceptibility of various cancer cell types to anticancer drugs. Here, we systematically investigated the CD44-dependent chemoresistance of NSCLC cells and developed a liposomal siRNA delivery system to overcome this chemoresistance by the self-targeted downregulation of CD44.

Methods

We confirmed the relationship between the expression of CD44 and the chemosensitivity of NSCLC cells using flow cytometry and MTT assay. We then generated and characterized cisplatin-resistant cell lines and compared the expression of CD44 in resistant cells to that in parental cells using western blotting. To evaluate whether the chemosensitivity of resistant cells depends on CD44 expression, we performed CD44 knockdown using CD44 siRNA and detected the chemosensitivity of these cells. Additionally, we prepared hyaluronic acid (HA)-coated liposomes as a targeted delivery system to selectively deliver CD44-specific siRNA to chemoresistant NSCLC cells and observed whether the chemosensitivity of these cells was improved.

Results

We found that CD44 expression is inversely proportional to the degree of cellular response to cisplatin chemotherapy and that CD44 is overexpressed in chemoresistant NSCLC cells. By performing CD44 knockdown using siRNA, we reconfirmed that the chemosensitivity of resistant cells depends on CD44 expression. We also observed that HA-liposome-mediated siRNA delivery prior to cisplatin chemotherapy significantly reduced CD44 expression and enhanced cisplatin sensitivity in chemoresistant NSCLC cells.

Conclusions

These results suggest that self-targeted downregulation of chemoresistance-associated cell surface proteins during chemotherapy is an effective therapeutic strategy for overcoming the chemoresistance of NSCLC cells.

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Data Availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

NSCLC:

Non-small cell lung cancer

HA:

Hyaluronic acid

CSC:

Cancer stem cell

MTT:

3-[4,5-Dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide

SF:

Surviving fraction

DiI:

1,1′-Dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate

BAX:

BCL2-associated X apoptosis regulator

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

DAPI:

2-(4-Amidinophenyl)-1H-indole-6-carboxamidine

MDR1:

Multidrug-resistant protein

MRP2:

Multidrug resistance-associated protein 2

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Funding

This work was supported by a Korea University Grant and the National Research Foundation of Korea (NRF) grant funded by the Ministry of Education, Science, and Technology (NRF-2017R1E1A1A01074847) and Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare (Grant no. HI17C0654).

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Authors and Affiliations

  1. Department of Thoracic and Cardiovascular Surgery, Korea University Guro Hospital, College of Medicine, Korea University, 148, Gurodong-ro, Guro-gu, Seoul, 08308, Republic of Korea

    Yu Hua Quan, Byeong Hyeon Choi, Young Ho Choi & Hyun Koo Kim

  2. Department of Biomedical Sciences, College of Medicine, Korea University, Seoul, 02841, Republic of Korea

    Yu Hua Quan, Byeong Hyeon Choi & Hyun Koo Kim

  3. Department of Bio and Brain Engineering and KAIST Institute for Health Science and Technology, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea

    Ji-Young Lim & Ji-Ho Park

  4. Department of Bio-Convergence, Korea University, Seoul, 02841, Republic of Korea

    Yeonho Choi

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  1. Yu Hua Quan
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Contributions

Conception and design: YHQ, J-HP, and HKK. Development of methodology: YHQ. Acquisition of data (provided materials, provided facilities, etc.): YHn, JL, and YHC. Analysis and interpretation of data (e.g. statistical analysis, biostatistics, computational analysis): YHQ. Writing, review, and/or revision of the manuscript: YHQ, J-HP, and HKK. Administrative, technical, or material support (i.e. reporting or organizing data, constructing databases): JL, YC, and YHC. Study supervision: J-HP and HKK.

Corresponding authors

Correspondence to Ji-Ho Park or Hyun Koo Kim.

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All authors read and approved the final version of the manuscript, and the authors declare no conflicts of interest.

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Supplementary material 1 (DOCX 13 KB)

Supplementary Fig. 1. Preparation of HA-liposomes for CD44-targeted siRNA delivery

. (A) The hydrodynamic sizes of 5- and 700-kDa HA-liposomes were approximately 144.2 and 156.4 nm, respectively. (B) The 700-kDa HA-liposomes showed notably higher targeting towards CD44-overexpressing H1299 and H1793 cells than H522 and H2087 cells which expressed low levels of CD44. (C) The 1:1 ratio of siRNA to protamine is optimized size for encapsulation of HA-liposomes. (D) Scheme of procedure to prepare siRNA-loaded HA-liposomes. (DOCX 178 KB)

Supplementary material 3 (DOCX 22 KB)

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Quan, Y.H., Lim, JY., Choi, B.H. et al. Self-targeted knockdown of CD44 improves cisplatin sensitivity of chemoresistant non-small cell lung cancer cells. Cancer Chemother Pharmacol 83, 399–410 (2019). https://doi.org/10.1007/s00280-018-3737-y

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