scholarly journals Purification and In Vitro Evaluation of an Anti-HER2 Affibody-Monomethyl Auristatin E Conjugate in HER2-Positive Cancer Cells

Biology ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 758
Author(s):  
Isabella Damiani ◽  
Silvia Castiglioni ◽  
Alicja Sochaj-Gregorczyk ◽  
Fabrizia Bonacina ◽  
Irma Colombo ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cancer Cells ◽  
Cell Lines ◽  
Triple Negative ◽  
Cancer Cell Line ◽  
Her2 Positive ◽  
Antimitotic Agent ◽  
High Affinity ◽  
Cancer Targeted Therapy

A promising approach for the development of high-affinity tumor targeting ADCs is the use of engineered protein drugs, such as affibody molecules, which represent a valuable alternative to monoclonal antibodies (mAbs) in cancer-targeted therapy. We developed a method for a more efficient purification of the ZHER2:2891DCS affibody conjugated with the cytotoxic antimitotic agent auristatin E (MMAE), and its efficacy was tested in vitro on cell viability, proliferation, migration, and apoptosis. The effects of ZHER2:2891DCS-MMAE were compared with the clinically approved monoclonal antibody trastuzumab (Herceptin®). To demonstrate that ZHER2:2891DCS-MMAE can selectively target HER2 overexpressing tumor cells, we used three different cell lines: the human adenocarcinoma cell lines SK-BR-3 and ZR-75-1, both overexpressing HER2, and the triple-negative breast cancer cell line MDA-MB-231. MTT assay showed that ZHER2:2891DCS-MMAE induces a significant time-dependent toxic effect in SK-BR-3 cells. A 30% reduction of cell viability was already found after 10 min exposure at a concentration of 7 nM (IC50 of 80.2 nM). On the contrary, MDA-MB-231 cells, which express basal levels of HER2, were not affected by the conjugate. The cytotoxic effect of the ZHER2:2891DCS-MMAE was confirmed by measuring apoptosis by flow cytometry. In SK-BR-3 cells, increasing concentrations of conjugated affibody induced cell death starting from 10 min of treatment, with the strongest effect observed after 48 h. Overall, these results demonstrate that the ADC, formed by the anti-HER2 affibody conjugated to monomethyl auristatin E, efficiently interacts with high affinity with HER2 positive cancer cells in vitro, allowing the selective and specific delivery of the cytotoxic payload.

Small Molecular Leads Differentially Active Against HER2 Positive and Triple Negative Breast Cancer Cell Lines

2019 ◽  
Vol 15 (7) ◽  
pp. 738-742 ◽  
Author(s):  
Adnan Badran ◽  
Atia-tul-Wahab ◽  
Sharmeen Fayyaz ◽  
Elias Baydoun ◽  
Muhammad Iqbal Choudhary
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Triple Negative ◽  
Cancer Cell Lines ◽  
Breast Cancer Cell Lines ◽  
Cancer Type

Background:Breast cancer is the most prevalent cancer type in women globally. It is characterized by distinct subtypes depending on different gene expression patterns. Oncogene HER2 is expressed on the surface of cell and is responsible for cell growth regulation. Increase in HER2 receptor protein due to gene amplification, results in aggressive growth, and high metastasis in cancer cells.Methods:The current study evaluates and compares the anti-breast cancer effect of commercially available compounds against HER2 overexpressing BT-474, and triple negative MDA-MB-231 breast cancer cell lines.Results:Preliminary in vitro cell viability assays on these cell lines identified 6 lead molecules active against breast cancer. Convallatoxin (4), a steroidal lactone glycoside, showed the most potent activity with IC50 values of 0.63 ± 0.56, and 0.69 ± 0.59 µM against BT-474 and MDA-MB-231, respectively, whereas 4-[4-(Trifluoromethyl)-phenoxy] phenol (3) a phenol derivative, and Reserpine (5) an indole alkaloid selectively inhibited the growth of BT-474, and MDA-MB-231 breast cancer cells, respectively.Conclusion:These results exhibited the potential of small molecules in the treatment of HER2 amplified and triple negative breast cancers in vitro.

scholarly journals Cytotoxic, genotoxic and apoptotic effects of Viburnum opulus on colon cancer cells: an in vitro study

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Kubra Bozali ◽  
Eray Metin Guler ◽  
Ahmet Sadik Gulgec ◽  
Abdurrahim Kocyigit
Keyword(s):  
Colorectal Cancer ◽  
Dna Damage ◽  
Cell Viability ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Methanolic Extract ◽  
Cancer Cell Line ◽  
Dependent Manner ◽  
Apoptotic Effects

AbstractObjectiveIntake of various fruits is quite significant for maintaining the human body, due to their supply of useful constituents. V. opulus has been found to have outstanding antioxidant activity while showing a pro-oxidant effect at high doses. Due to this feature, V. opulus would be anticipated to have a healing impact on cancer treatment. In this study, it has been proposed to examine the cytotoxic, genotoxic, and apoptotic effects of V. opulus on human colorectal cancer cell.MethodDifferent concentrations of V. opulus methanolic extract (5–2000 μg/mL) were incubated for 24 h with colorectal cancer cell line (Lovo). The cell viability, intracellular reactive oxygen species (iROS), DNA damage, and apoptosis were measured after incubation.ResultsThe obtained results of this research demonstrate decreased cell viability and increased DNA damage, iROS, and apoptosis levels of V. opulus in Lovo cells in a concentration-dependent manner in the range of 14.88–52.06%. There were strong positive relationships between apoptosis, genotoxicity, and cytotoxicity in V. opulus methanolic extract treated cancer cell line.DiscussionThis in vitro research clearly demonstrated that V. opulus methanolic extract induces DNA damage, apoptosis, and cytotoxicity in a dose-dependent manner in cancer cells due to its pro-oxidant activity.ConclusionAlthough in vitro results are favorable, in vivo and further studies are needed.

scholarly journals Enhanced Antitumor Effect of Trastuzumab and Duligotuzumab or Ipatasertib Combination in HER-2 Positive Gastric Cancer Cells

Cancers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 2339
Author(s):  
Maria Maddalena Laterza ◽  
Vincenza Ciaramella ◽  
Bianca Arianna Facchini ◽  
Elisena Franzese ◽  
Carmela Liguori ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Antitumor Effect ◽  
Combined Treatment ◽  
Human Gastric Cancer ◽  
Synergistic Activity ◽  
Her2 Positive ◽  
Gastric Cancer Cells

The anti-HER2 monoclonal antibody trastuzumab is a key drug for the treatment of HER2-positive gastric cancer (GC); however, its activity is often limited by the onset of resistance and mechanisms of resistance are still poorly understood. Several targeted agents showed synergistic activity by concomitant use with trastuzumab in vitro and are under clinical investigation. The aim of this study was to assess the antitumor activity of duligotuzumab, an anti HER3/EGFR antibody or ipatasertib, an AKT inhibitor, combined with trastuzumab in a panel of HER2-positive human gastric cancer cells (GCC), and the efficacy of such combinations in HER2-resistant cells. We have assessed the efficacy of duligotuzumab or ipatasertib and trastuzumab in combination, analyzing proliferation, migration and apoptosis and downstream intracellular signaling in vitro on human HER2-positive GCC (NCI-N87, OE33, OE19) and in negative HER2 GCC (MKN28). We observed a reduction of proliferation, migration and apoptotic rate in HER2-positive OE33, OE19 and N87 cell lines with the combination of duligotuzumab or ipatasertib plus trastuzumab. In particular, in OE33 and OE19 cell lines, the same combined treatment inhibited the activation of proteins downstream of HER2, HER3, AKT and MAPK pathways. Targeting both HER2 and HER3, or HER2 and AKT, results in an improved antitumor effect on HER2-positive GCC.

scholarly journals Efficacy and mechanism of KRAS G12C PROTAC in inhibiting the proliferation of pancreatic cancer cells

2022 ◽  
Vol 3 (1) ◽  
pp. 176-184
Author(s):  
Shuai Gao ◽  
◽  
Fangxia Zou ◽  
Lixia Zheng ◽  
Yunjie Wang ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Pancreatic Cancer ◽  
Cell Viability ◽  
Cancer Cells ◽  
Pancreatic Cancer Cell Line ◽  
Cancer Cell Line ◽  
26S Proteasome ◽  
Mechanistic Study ◽  
Pancreatic Cancer Cells

Pancreatic cancer is a rare but highly malignant cancer with few effective treatments available. Targeting cancers bearing specific genetic mutations offers a new approach for cancer therapy. PROTAC (proteolysis-targeting chimeras) is an emerging technique to design targeted therapy and increasing evidence supports its utility. This study examined the in vitro pharmacodynamics and mechanism of PROTAC K-Ras Degrader-1 (PKD-1), a PROTAC molecule, in inhibiting the proliferation of pancreatic cancer cells. We used a pancreatic cancer cell line, MIA PaCa-2 cells, to examined the binding and degradation-promoting capabilities of PKD-1 on KRAS G12C protein and further evaluated the effects of PKD-1 on cell viability, cell cycle and apoptosis. PKD-1 was able to bind to KRAS G12C protein, promoted its degradation for up to 72 h, reduced cell viability, increased cell cycle arrest and promoted cell apoptosis. Mechanistic study found that the efficacy of PKD-1 was at least partially mediated by promoting 26S proteasome degradation process. Combined, these results extended previous findings and support the potential utility of PROTAC molecules such as PKD-1 as a new treatment strategy against pancreatic cancer.

scholarly journals An Investigation on the in vitro Effects of Elaidic Acid and Gamma-Linolenic Acid on the Survival of MDA-MB-231 and RKO Cell Lines

2020 ◽  
Vol 2 (1) ◽  
pp. 12
Keyword(s):  
Breast Cancer ◽  
Colon Cancer ◽  
Cell Viability ◽  
Cell Lines ◽  
Cancer Cell ◽  
Statistical Difference ◽  
Cancer Cell Line ◽  
Colon Cancer Cell ◽  
Gamma Linolenic Acid

Colorectal and breast cancer are a major cause of mortality worldwide. They can be caused due to an array of factors, of which diet plays an important role. Previous studies have suggested that Elaidic acid (EA), a trans fatty acid, supports the growth of colon cancer cell lines, like-RKO, LoVo, and HT29;. In contrast, Gamma-linolenic acid (GLA), a poly-unsaturated fatty acid, has tumoricidal effects. Therefore, the objective of this study was to investigate and delineate the in vitro dose-dependent effects of EA and GLA on the survival of MDA-MB-231(breast cancer cell line) and RKO cells (colon cancer cell line), through cell viability assay. The principal findings of this study were that EA induced a stimulatory effect. At the same time, GLA had an inhibitory effect on both cell lines. There was no statistical significance in the percentage viability of the MDA-MB-231 cells after treatment with EA. At the same time, there was a statistical difference in percentage viability after treatment with GLA. The highest test concentration of GLA that caused approximately 99% inhibition of MDA-MB-231 cells was 500µM, and its IC50 was mathematically calculated to be 239.687µM. For RKO cells, there was a statistical difference in percentage cell viability after treatment with EA. At the same time, there was no statistical difference in percentage cell viability after treatment with GLA. These results suggested that MDA-MB-231 cells were more susceptible to the effects of EA and GLA and also that EA should be consumed in moderation, while GLA appeared as a promising therapeutic option either by itself or in combination with other chemotherapeutic drugs.

scholarly journals In vitro Anticancer Activity of Marine Sponges Against T47D and HeLa Cell Lines

2020 ◽  
Vol 19 (1) ◽  
pp. 25-28
Author(s):  
Suciati ◽  
Lusiana Arifianti
Keyword(s):  
Anticancer Activity ◽  
Cancer Cells ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Anticancer Agents ◽  
Cancer Cell Line ◽  
Marine Sponges ◽  
Hela Cell Lines

Marine sponges have been known as the source of natural products. Various metabolites with potent bioactivities have been reported from this organism. The current study aims to investigate the anticancer potency of three marine sponges namely Diacarnus debeauforti, Haliclona amboinensis and Agelas cavernosa collected from Barrang Lompo Island, South Sulawesi, Indonesia. The ethyl acetate extracts of the sponges were screened against T47D breast cancer cells and HeLa cervical cancer cells by using the MTT method. The results showed that these sponges demonstrated anticancer activity against both cancer cell lines. The lowest IC50 of 18.2 μg/ml was given by the extract of A. cavernosa against T47D cell line, while in the screening against HeLa cancer cell line, the extract of D. debeauforti revealed the highest potency with IC50 of 15.7 μg/ml. Our results suggested that the marine sponges namely D. debeauforti, H. amboinensis and A. cavernosa can be good candidates for the development of anticancer agents. Dhaka Univ. J. Pharm. Sci. 19(1): 25-28, 2020 (June)

scholarly journals Multi-Omics Investigation of Innate Navitoclax Resistance in Triple-Negative Breast Cancer Cells

Cancers ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 2551
Author(s):  
Michal Marczyk ◽  
Gauri A. Patwardhan ◽  
Jun Zhao ◽  
Rihao Qu ◽  
Xiaotong Li ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Recovery Period ◽  
Genome Structure ◽  
Copy Number Variations ◽  
Drug Induced

Cancer cells employ various defense mechanisms against drug-induced cell death. Investigating multi-omics landscapes of cancer cells before and after treatment can reveal resistance mechanisms and inform new therapeutic strategies. We assessed the effects of navitoclax, a BCL2 family inhibitor, on the transcriptome, methylome, chromatin structure, and copy number variations of MDA-MB-231 triple-negative breast cancer (TNBC) cells. Cells were sampled before treatment, at 72 h of exposure, and after 10-day drug-free recovery from treatment. We observed transient alterations in the expression of stress response genes that were accompanied by corresponding changes in chromatin accessibility. Most of these changes returned to baseline after the recovery period. We also detected lasting alterations in methylation states and genome structure that suggest permanent changes in cell population composition. Using single-cell analyses, we identified 2350 genes significantly upregulated in navitoclax-resistant cells and derived an 18-gene navitoclax resistance signature. We assessed the navitoclax-response-predictive function of this signature in four additional TNBC cell lines in vitro and in silico in 619 cell lines treated with 251 different drugs. We observed a drug-specific predictive value in both experiments, suggesting that this signature could help guiding clinical biomarker studies involving navitoclax.

scholarly journals COMPARATIVE IN-VITRO ANTICANCER ACTIVITY OF THANGA PARPAM (SIDDHA GOLD DRUG) IN BREAST, LIVER, PROSTATE AND LUNG CANCER CELL LINES

2021 ◽  
Vol 12 (1) ◽  
pp. 64-67
Author(s):  
Sulthan Shajahan ◽  
Arul Amuthan
Keyword(s):  
Prostate Cancer ◽  
Lung Cancer ◽  
Particle Size ◽  
Cell Viability ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines ◽  
Prostate Cancer Cells

Thanga parpam is a gold nanoparticle used in Siddha for many chronic and challenging diseases including cancers. Clinically it shows benefit in some cancer types, but not to all types. This study was aimed to compare the in-vitro anticancer activity of Thanga parpam in various cancer cell lines. The particle size and elements were evaluated using Scanning Electron Microscope coupled with Energy Dispersive X-Ray Spectroscopy. Thanga parpam was added to breast adenocarcinoma cells, Human hepatocellular carcinoma cells, Human prostate cancer cells and Human lung adenocarcinoma epithelial cells for 24 hours. MTT assay was used to evaluate the cell viability. Graph was drawn from the % cell viability and dose required to kill 50 % cells was calculated. The gold particles are irregular disk shaped, but agglomerated to each other and not in uniform size. The particle size varies from 17.8 nm – 448 nm. About 9.3% of gold element was present in oxide and sulfide form. It showed dose dependent killing effect on all cancer cell lines. IC50% value was 0.63, 3.51, 6.65 and 11.01 µg/ml for breast, liver, prostate and lung cancer cells respectively. Thanga parpam is a potent anticancer drug to all the four cancer cells, however higher efficacy was observed in breast, liver and prostate cancer cells.

Stew in its Own Juice: Protein Homeostasis Machinery Inhibition Reduces Cell Viability in Multiple Myeloma Cell Lines

2019 ◽  
Vol 19 (2) ◽  
pp. 112-119 ◽  
Author(s):  
Mariana B. de Oliveira ◽  
Luiz F.G. Sanson ◽  
Angela I.P. Eugenio ◽  
Rebecca S.S. Barbosa-Dantas ◽  
Gisele W.B. Colleoni
Keyword(s):  
Multiple Myeloma ◽  
Cell Death ◽  
Cell Viability ◽  
Cell Lines ◽  
Treatment Options ◽  
Compensatory Mechanism ◽  
Protein Homeostasis ◽  
Protein Response ◽  
Rpmi 8226

Introduction:Multiple myeloma (MM) cells accumulate in the bone marrow and produce enormous quantities of immunoglobulins, causing endoplasmatic reticulum stress and activation of protein handling machinery, such as heat shock protein response, autophagy and unfolded protein response (UPR).Methods:We evaluated cell lines viability after treatment with bortezomib (B) in combination with HSP70 (VER-15508) and autophagy (SBI-0206965) or UPR (STF- 083010) inhibitors.Results:For RPMI-8226, after 72 hours of treatment with B+VER+STF or B+VER+SBI, we observed 15% of viable cells, but treatment with B alone was better (90% of cell death). For U266, treatment with B+VER+STF or with B+VER+SBI for 72 hours resulted in 20% of cell viability and both treatments were better than treatment with B alone (40% of cell death). After both triplet combinations, RPMI-8226 and U266 presented the overexpression of XBP-1 UPR protein, suggesting that it is acting as a compensatory mechanism, in an attempt of the cell to handle the otherwise lethal large amount of immunoglobulin overload.Conclusion:Our in vitro results provide additional evidence that combinations of protein homeostasis inhibitors might be explored as treatment options for MM.

A Novel Triazole Derivative Drug Presenting In Vitro and In Vivo Anticancer Properties

2018 ◽  
Vol 18 (17) ◽  
pp. 1483-1493
Author(s):  
Ricardo Imbroisi Filho ◽  
Daniel T.G. Gonzaga ◽  
Thainá M. Demaria ◽  
João G.B. Leandro ◽  
Dora C.S. Costa ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cell Line ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Hepatic Function ◽  
Anticancer Properties ◽  
Mcf 7

Background: Cancer is a major cause of death worldwide, despite many different drugs available to treat the disease. This high mortality rate is largely due to the complexity of the disease, which results from several genetic and epigenetic changes. Therefore, researchers are constantly searching for novel drugs that can target different and multiple aspects of cancer. Experimental: After a screening, we selected one novel molecule, out of ninety-four triazole derivatives, that strongly affects the viability and proliferation of the human breast cancer cell line MCF-7, with minimal effects on non-cancer cells. The drug, named DAN94, induced a dose-dependent decrease in MCF-7 cells viability, with an IC50 of 3.2 ± 0.2 µM. Additionally, DAN94 interfered with mitochondria metabolism promoting reactive oxygen species production, triggering apoptosis and arresting the cancer cells on G1/G0 phase of cell cycle, inhibiting cell proliferation. These effects are not observed when the drug was tested in the non-cancer cell line MCF10A. Using a mouse model with xenograft tumor implants, the drug preventing tumor growth presented no toxicity for the animal and without altering biochemical markers of hepatic function. Results and Conclusion: The novel drug DAN94 is selective for cancer cells, targeting the mitochondrial metabolism, which culminates in the cancer cell death. In the end, DAN94 has been shown to be a promising drug for controlling breast cancer with minimal undesirable effects.

Sign in / Sign up

Export Citation Format

Share Document