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Helas3

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HeLaS3 is a cell line derived from human cervical cancer cells. It is a subclone of the HeLa cell line, which is one of the most widely used and well-characterized human cell lines in biomedical research. The HeLaS3 cell line is commonly used in various laboratory experiments and assays.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Mcf 7, by RIKEN BioResource Center (1 mentions) Nci h460, by RIKEN BioResource Center (1 mentions) Hepg2, by RIKEN BioResource Center (1 mentions) Bezafibrate, by Fujifilm (1 mentions) Hek293, by American Type Culture Collection (1 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Hek293t, by American Type Culture Collection (1 mentions)

9 protocols using helas3

1

Cancer Cell Line Cultivation and Maintenance

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The human cancer cell lines HeLaS3, KBvin, MCF-7, HepG2, NCI-H460 and NCI-H460/MX20 were grown in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) and 1/100 (v/v) penicillin/streptomycin at 37 °C under 5% carbon dioxide. The medium was routinely changed every two days. As cell confluence reached 80% in the medium, the cells were treated with 0.25% of trypsin-EDTA for the next passage. Human cervical carcinoma cell line HeLaS3, human non-small-cell lung carcinoma cell line NCI-H460, human liver carcinoma cell line HepG2 and human breast cancer cell line MCF7 were purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan). The multidrug-resistant human cervical cancer cell line KB-vin was kindly provided by Dr. Kuo-Hsiung Lee (University of North Carolina, Chapel Hill, NC, USA) and maintained with 100 nM of vincristine each week. MDR human non-small-cell lung carcinoma cell line NCI-H460/MX20 was the resistant cell line selected from NCI-H460. It was treated with gradually increasing concentrations of mitoxantrone and maintained in 20 nM of mitoxantrone.
Note: all tested compounds were dissolved in DMSO and the final concentrations of DMSO in all experiments were less than 0.1%.
Chen C.Y., Lien J.C., Chen C.Y., Hung C.C, & Lin H.C. (2021). Design, Synthesis and Evaluation of Novel Derivatives of Curcuminoids with Cytotoxicity. International Journal of Molecular Sciences, 22(22), 12171.
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2

Establishing P-gp-expressing and multi-drug resistant cancer cell lines

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Human stably P-gp-expressing cells (ABCB1/Flp-InTM-293) were established and cultured in DMEM as in a previous study [27 (link)]. The human cervical epithelioid carcinoma cell line HeLaS3 was purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan), and the multi-drug resistant human cervical cancer cell line KBvin was kindly provided by Dr. Kuo-Hsiung Lee (University of North Carolina, Chapel Hill, NC, USA). All cancer cell lines were cultured in RPMI-1640 containing 10% FBS, at 37 °C in a humidified atmosphere of 5% CO2.
Hung H.Y., Hung C.C., Liang J.W., Chen C.F., Chen H.Y., Shieh P.C., Kuo P.C, & Wu T.S. (2019). Constituents and Anti-Multidrug Resistance Activity of Taiwanofungus camphoratus on Human Cervical Cancer Cells. Molecules, 24(20), 3730.
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3

Culturing Human Cancer Cell Lines

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Human cervical epithelioid carcinoma HeLaS3 was purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan), and the multi-drug resistant human cervical cancer cell line KB/VIN was kindly provided by Dr. Kuo-Hsiung Lee (University of North Carolina, Chapel Hill, NC, USA) and maintained with vincristine regularly. The human P-gp stable expression cells (ABCB1/Flp-InTM-293) and parental cell line Flp-InTM-293 were constructed as previously described [44 (link)]. All cells were cultured in DMEM or RPMI-1640 containing 10% FBS at 37 °C in a humidified atmosphere of 5% CO2.
Teng Y.N., Wang C.C., Liao W.C., Lan Y.H, & Hung C.C. (2020). Caffeic Acid Attenuates Multi-Drug Resistance in Cancer Cells by Inhibiting Efflux Function of Human P-Glycoprotein. Molecules, 25(2), 247.
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4

Cell Culture of Cervical Cancer Lines

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Human cervical carcinoma cell line HeLaS3 was purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan). The KB-vin was multidrug resistant human cervical cancer cell line, was kindly gave from Dr. Kuo-Hsiung Lee (University of North Carolina, Chapel Hill, NC, USA). All cells were cultured in RPMI-1640 supplemented with 10% FBS at 37 °C in a humidified 5% CO2 incubator.
Chen H.J., Chung Y.L., Li C.Y., Chang Y.T., Wang C.C., Lee H.Y., Lin H.Y, & Hung C.C. (2018). Taxifolin Resensitizes Multidrug Resistance Cancer Cells via Uncompetitive Inhibition of P-Glycoprotein Function. Molecules, 23(12), 3055.
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5

Cell Culture Maintenance and Compound Usage

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Cells were maintained in the following culture medium: C2C12 (mouse myoblast, American Type Culture Collection [ATCC], CRL-1772), HeLa S3 (RIKEN BioResource Research Center, RCB1525), HEK293 (human embryonic kidney, ATCC, CRL-1573), HEK293T (ATCC, CRL-3216), A375 (human malignant melanoma, ATCC, CRL-1619), and H2009 (human lung adenocarcinoma, ATCC, CRL-5911) cells were maintained in DMEM, high glucose, GlutaMAX supplement (Thermo Fisher Scientific, 10566016) with 10% fetal bovine serum (FBS); H2122 (human lung adenocarcinoma, ATCC, CRL-5985), H1944 (human lung adenocarcinoma, ATCC, CRL-5907), and H441 (human lung adenocarcinoma, ATCC, HTB-174) cells were maintained in RPMI 1640 medium (Thermo Fisher Scientific, A1049101) with 10% FBS and 1% penicillin-streptomycin (Thermo Fisher Scientific, 15140148). All cells were cultured in a humidified incubator under 5% CO2 at 37°C.
The following compounds were used in this study: ANS (Chem-Impex International), CAP (FUJIFILM Wako Pure Chemical Corporation), CHX (Sigma-Aldrich), EME (Cayman Chemical), and bezafibrate (FUJIFILM Wako Pure Chemical Corporation).
Kimura Y., Saito H., Osaki T., Ikegami Y., Wakigawa T., Ikeuchi Y, & Iwasaki S. (2022). Mito-FUNCAT-FACS reveals cellular heterogeneity in mitochondrial translation. RNA, 28(6), 895-904.
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6

Characterizing Cell Line Drug Resistance

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Drug-sensitive HeLa S3 and KB cancer cell lines were from Bioresource Collection and Research Center (Hsinchu, Taiwan). The KBvin cell line was a generous gift from Dr. Kuo-Hsiung Lee (University of North Carolina, Chapel Hill, NC, USA). The parental cells Flp-InTM-293 were incubated with 100 μg/mL zeocin. ABCB1-expressing cells (ABCB1/Flp-InTM-293) with the stable expression of human P-gp were cultured with 100 μg/mL hygromycin B. The Flp-InTM-293 and ABCB1/Flp-InTM-293 cell lines were established as described previously [29 (link)]. A total of five cell lines were incubated in DMEM or RPMI 1640 with 10% FBS and incubated at 37 °C under 5% CO2.
Wu I.T., Kuo C.Y., Su C.H., Lan Y.H, & Hung C.C. (2023). Pinostrobin and Tectochrysin Conquer Multidrug-Resistant Cancer Cells via Inhibiting P-Glycoprotein ATPase. Pharmaceuticals, 16(2), 205.
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7

Multidrug Resistant Cancer Cell Lines

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The multidrug resistant cancer cell line, KB-vin, was a generous gift from Dr. Kuo-Hsiung Lee (University of North Carolina, Chapel Hill, U.S.A) and maintained with vincristine. The drug sensitive cell line, HeLaS3, was purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan). Human endometrial epithelial cells (EEC) were purchased from Shanghai Institute of Cell Biology (Shanghai, China). All cells were cultured in RPMI-1640 containing 10% FBS at 37 °C in a humidified atmosphere of 5% CO2.
Ling Y., Yang Q.X., Teng Y.N., Chen S., Gao W.J., Guo J., Hsu P.L., Liu Y., Morris-Natschke S.L., Hung C.C, & Lee K.H. (2018). Development of novel amino-quinoline-5,8-dione derivatives as NAD(P)H:quinone oxidoreductase 1 (NQO1) inhibitors with potent antiproliferative activities. European journal of medicinal chemistry, 154, 199-209.
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8

Establishment of Cell Lines for MDR Studies

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The human P-gp stable expression cells (ABCB1/Flp-InTM-293) and parental cell line Flp-InTM-293 were constructed as previous described [17 (link)]. Human cervical epithelioid carcinoma HeLaS3 was purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan). The multi-drug-resistant human cervical cancer cell line KB/VIN was a generous gift from Dr. Kuo-Hsiung Lee (University of North Carolina, Chapel Hill, NC, USA). The resistance of KB/VIN was maintained with regular vincristine treatment. All cells were cultured in DMEM or RPMI-1640 containing 10% FBS at 37 °C in a humidified atmosphere of 5% CO2.
Teng Y.N., Wang Y.H., Wu T.S., Hung H.Y, & Hung C.C. (2019). Zhankuic Acids A, B and C from Taiwanofungus camphoratus Act as Cytotoxicity Enhancers by Regulating P-Glycoprotein in Multi-Drug Resistant Cancer Cells. Biomolecules, 9(12), 759.
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9

Culturing Human Cervical Cancer Cells

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Human cervical carcinoma cell line HeLaS3 was purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan). The multidrug resistant human cervical cancer cell line KB-vin was a generous gift from Dr. Kuo-Hsiung Lee (University of North Carolina, Chapel Hill, USA) and maintained with vincristine in a fixed period. All cells were cultured in RPMI-1640 containing 10% FBS at 37 °C in a humidified atmosphere of 5% CO2.
Chang Y.T., Wang C.C., Wang J.Y., Lee T.E., Cheng Y.Y., Morris-Natschke S.L., Lee K.H, & Hung C.C. (2018). Tenulin and isotenulin inhibit P-glycoprotein function and overcome multidrug resistance in cancer cells. Phytomedicine : international journal of phytotherapy and phytopharmacology, 53, 252-262.
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