The Dual Influence of Adrenaline may be Effective in the Treatment of Grade 3 and 4 Cancers with a Synergistic Effect with Chemotherapy

doi:10.21203/rs.3.rs-5313600/v1

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The Dual Influence of Adrenaline may be Effective in the Treatment of Grade 3 and 4 Cancers with a Synergistic Effect with Chemotherapy

https://doi.org/10.21203/rs.3.rs-5313600/v1

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Objective(s): The present study investigated the impact of physiological and pharmacological concentrations of epinephrine alone and together with a β-adrenergic receptor antagonist (propranolol) on the proliferation/migration rate, substrate-adhesion ability, and viability of two slowing-growing brain glioblastoma (U87) and fast-growing gastric adenocarcinoma (AGS) cell lines.

Materials and Methods: The cells were treated with epinephrine in a 16 µM (physiological) – 256 µM (pharmacological) concentration range and combined with 75 µM propranolol. The cell proliferation/migration rate was determined using scratch wound healing assay at 12 h and 24 h post-treatment. The substrate-adhesion ability was examined using a plate-and-wash assay after detaching the cells with EDTA/trypsin solution and incubating the cells with the drug treatments for 15 and 30 min. The cell viability was assessed using the trypan blue dye exclusion method with hemocytomer count.

Results: The results showed that epinephrine had a dual effect; enhancing the proliferation/migration, adhesion, and viability of both cells at physiological concentrations while suppressing the mentioned features at pharmacological concentrations. Propranolol served antagonistic activity, especially at the low doses of epinephrine.

Conclusion: The study suggests that combination therapy with epinephrine and propranolol can shift the cancer cell growth and metastasis-promoting features of epinephrine toward cancer cell eradication for both slowing-growing U87 and Fast-growing AGS cells.

Epinephrine

Gastric cancer

Brain tumor

Cancer

Glioblastoma

Evidence from human and animal studies has shown that chronic stress with prolonged activation of the sympathetic nervous system and secretion of the stress hormones (norepinephrine and epinephrine) may influence tumor initiation and progression (1). The correlation between tumor growth and metastasis with epinephrine secretion has been shown in multiple clinical and epidemiological studies. In both acute and chronic phases of stress, the catecholamines are elevated in the blood and bind to adrenergic receptors (2), which are widely expressed in various mammalian tissues.

The effects of catecholamine on target cells through β-adrenergic receptors have been shown in several cancers (4). Following stimulation of β-adrenergic receptors, the intracellular cAMP level alters in the cells affecting cell proliferation and differentiation (5). Blocking the β-adrenergic receptors using antagonists such as propranolol is accompanied by the reduction of tumor progression and metastasis in different mouse models of cancers such as breast, and prostate carcinomas, malignant melanoma, and leukemia (6). Norepinephrine also affects epithelial-mesenchymal transformation as a crucial process in tumor metastasis and invasion in different malignancies including prostate, ovary, gastric, colorectal, and lung cancer (7, 8, 9, 10, 11). On the other hand, norepinephrine through β2-adrenergic receptors decreased breast cancer cell invasion. The complexity of the β2-adrenergic receptors signaling pathway may play a role in this unexpected result (7). Nevertheless, it has been proven that catecholamine and their associated catecholamine are involved in different cancers.

Gastric cancer is the third cause of death of all malignancies and also the 5th most common neoplasm worldwide (8). It has been estimated that adenocarcinomas are the most prevalent type of all malignant gastric tumors (about 95%) and the remaining %5 comprise lymphomas, stromal tumors, and other rare tumors (12). The clinical outcomes of gastric cancers remain poor and many cases are diagnosed in late stages, due to a combination of genetic and environmental factors (13). It is shown that there is a positive correlation between stress/ depression and gastric cancer (14). This indicates that stress hormones and their associated receptors might also be involved in gastric cancer.

Similarly, glioblastoma multiform (GBM), the fourth stage of astrocytoma, is the most common primary malignant tumor of the central nervous system (CNS) that occurs in 1/10,000 patients (15). As with gastric cancer, this type of cancer has a poor prognosis and is recalcitrant to contemporary therapies, i.e. combination therapies with surgery, chemo- and radiotherapy (16). The disease is severely intolerable, and patients generally give up within 14 months after diagnosis (17). Studies illustrated that β-adrenergic receptors (β-AR) are closely associated with the occurrence and development of brain tumors. β-AR agonists might contribute to promoting glioblastoma cell line (U251) proliferation and migration (U251) via phosphorylation of extracellular signal-regulated kinase 1/2 (ERK ½) and overexpression of matrix metalloproteinases (MMPs), and the β-ARs antagonist (propranolol) might block this process.

Concerning the involvement of catecholamines and β-AR in cancer proliferation and metastasis, The present study attempts to investigate the effect of epinephrine through the β-adrenergic receptor and its antagonist (propranolol) on the proliferation, viability, and adhesion of AGS cell line of gastric adenocarcinoma as one of the most prevalent neoplasms in the world and U87 cell line of glioblastoma as one of rare cancer and the most aggressive malignant primary brain tumor in the world (18).

Materials

Epinephrine bitartrate (Product No. E1016. Lot No: 124K8803) and propranolol hydrochloride (CAS number, 318-98-9) were obtained from Sigma-Aldrich, USA, and maintained at 2-8° C. All cell-related reagents were purchased from Gibco, USA, including fetal bovine serum (FBS), L-glutamine, trypsin–EDTA solution, and trypan blue dye.

Cell growing conditions

The AGS human gastric adenocarcinoma and U87 glioblastoma cell lines were obtained from the Iranian Biological Resource Centre (Tehran, Iran). Both cell lines were cultured in RPMI-1640 medium supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS), 2 mM L-glutamine, as well as 100 IU/ml penicillin and 100 mg/ml streptomycin at 37˚C in a humidified incubator with 5% CO₂ atmosphere. At ~80 to 90 % cell conﬂuency, the cells were harvested by 0.1 ml trypsin–EDTA solution trypsin and the cell viability was measured by trypan blue dye exclusion (16) before the experiment.

Treatment groups

Both cells were treated with a varied range (16, 64, 128, 256 μM) of epinephrine per se and in combination with propranolol at a dose of 75 μM for 12 h and 24 h. The doses of the drugs were chosen according to our previous study (19).

Scratch wound healing assay

The proliferation and migration capability of the treated cells were determined using the in vitro scratch wound assay (17). Accordingly, the cells were grown to confluence (90%) in a Falcon® 24-well flat-bottom TC-treated cell culture plate, and with a streak of the tip of a sterile pipette, a scratch was created on the well bottom covered with monolayer cells. The well was then washed with phosphate-buffered saline (1X, pH 7.4) to remove the detached cells, was subjected to the treatments, and monitored for the cell migration into the ‘wound’ area at 0, 12, and 24 h post-treatment under a microscope (Motic optical microscope, model AE 31, Spain). Lastly, the images taken at the intervals were examined by a blinded observer who scored the wound-healing capacity from 1 to 6 according to Table 1 (17).

Table1.Index of wound healing time

cell growth & repair rate	Index of wound healing time
No repair	1
Somewhat	2
Half	3
More than half	4
Nearly complete	5
Complete repair	6

Cell adhesion assay

The adhesion ability of the cells was monitored using a modified plate-and-wash assay. Hence, the percentage of adhering cells was assessed following 0.5 mM EDTA + 0.25% w/v trypsin and 2.5 mM EDTA + 1.25% w/v trypsin pre-treatment (for 5 min) for the treatment (drugs) groups. The detached cells treated with the two concentrations of the cell-detaching reagents were pulled together for each group of treatment after the removal of the reagents via excessive washup and centrifugation process and transferred along with the drugs to the fresh plates. The number of detached cells was examined after 15 and 30 minutes of rest and incubation in the incubator, the plate was washed to remove the non-adherent cells and resuspended in an equal volume of PBS. Subsequently, the number of detached cells was measured using the hemocytometer counting technique.

Cell viability

The cell viability was assessed using trypan blue dye exclusion for the treatment groups in percentage terms at 0, 12, and 24 h (18).

Statistical analysis

The statistical analysis was conducted using GraphPad Prism version 6 (GraphPad Software, San Diego, CA) at a significance threshold of < 0.05. Descriptive data were expressed as mean ± standard deviation (SD). One-way analysis of variances (ANOVA) was performed followed by Tukey's post-comparision.

Epinephrine reduces the cells’ proliferation/migration rate

Fig. 1 shows the images of the wound healing assay monitored for 24 h. As can be seen, both non-treated U87 and AGS cells gradually grew and repaired the wound incompletely during the 24-hour incubation period (the first column in both Fig. 1A and Fig. 1B, respectively). Epinephrine at 16 µM low concentration contributed to the complete repair of the wound in both U87 (the second column in Fig. 1A) and AGS cells (the second column in Fig. 1B). However, at an extremely high concentration of 256 µM, epinephrine was found to be highly toxic and eradicated the cells, especially the seemingly more sensitive U87 cells (the third column in Fig. 1A). Propranolol exhibited a counteracting activity as opposed to that of epinephrine high concentrations and protected the cells against the cytotoxic activity of high-dose epinephrine.

In line with Fig.1, Fig .2 illustrates the cell proliferation/migration (wound healing) rate of U87 and AGS cells following treatment of the ‘scratched wound’ with epinephrine at increasing concentrations per se and together with propranolol at 12- and 24-h incubation time. Data are expressed according to the Table 1 rated system. In general, epinephrine reduced the proliferation/migration rate of the cells in a dose-dependent manner, and propranolol counteracted the epinephrine’s effect.

At 12 post-treatment, epinephrine reduced the cell proliferation/migration rate of U87 cancer cells significantly, especially at extremely high concentrations of 128 μM and 256 μM (Fig. 2A). The epinephrine effect was by almost three rates lower than that of the control (P<0.001). Epinephrine exerted no effect on the wound healing process at 16 μM. Adding 75 μM propranolol to the wells, remarkably lifted the cell proliferation/migration rate.

Similar findings were observed with the AGS cells, but to a lesser extent at 12 h (Fig. 2B). At most, epinephrine reduced the cell proliferation/migration by one rate at only 256 μM concentration (P<0.05). Again, propranolol recovered the cell proliferation/migration rate fully.

However, increasing the incubation time from 12 h to 24 h pronounced the impact of epinephrine (Fig. 2C & D). Epinephrine remarkedly decreased the cell proliferation/migration rate in both U87 (Fig. 2C) and AGS cell-grown wells (Fig. 2D). It almost completely hindered the wound healing rate at high 126 and 256 μM concentrations in both U87 and AGS cells (P<0.0001). Although propranolol limited the wound healing rate again at 24 h, its restricting effect was not as significant as it was in the wells treated for 12 h.

Epinephrine increases the number of detached cells

Fig. 3 shows the number of detached cells following incubation with the treatments. Data are expressed in terms of number of cells/ml withdrawn from the plates. Generally, epinephrine increased the number of detached cells/ml in a dose-dependent manner, while the propranolol addition reduced their numbers.

At 15 min incubation time, the number of detached cells increased remarkably with the increase in the epinephrine concentration in both U87 (Fig. 3A) and AGS cells (Fig 3B). At the highest concentration, epinephrine elevated the number of detached U87 cells by nearly 3-fold (Fig. 3A, P<0.0001). Epinephrine displayed no alteration in the number of detached cells at 16 μM concentration as compared to the control untreated group. Co-treatment with propranolol caused a nonsignificant reduction in the number of detached cells for all combinations. Epinephrine also elevated the number of detached AGS cells but to a lesser extent (Fig. 3B, P<0.01). It doubled the number. Likewise, the addition of propranolol also caused a nonsignificant decrease in the number of detached cells for all epinephrine doses.

The same pattern is also reflected in the 30-minute drug treatment of the cells (Fig. 3C & D). Epinephrine gradually enhanced the number of detached cells with increasing concentration, reaching approximately to three-fold increase in the number of detached U78 cells at the highest 256 μM concentration as opposed to the control (Fig. 3C, P< 0.001). Again, 16 μM epinephrine dose was insufficient to make a change in the number of detached cells. However, this time, the addition of 75 µM propranolol brought about a significant reduction in the number of detached cells, especially for the epinephrine/propranolol combination (256 µM/75 µM) as opposed to the counterpart single-agent treatment, i.e. epinephrine (256 µM, P < 0.001). A similar outcome was found for AGS cells (Fig 3D); epinephrine increased the number of detached cells in a dose-dependent and significant manner, and propranolol reduced significantly the number of detached cells in combination with 256 µM epinephrine in comparison with 256 µM epinephrine alone (P<0.001). Outstandingly, in this type of cell and at 30 min incubation time, 16 µM epinephrine seemed enough to make a significant change in the number of detached cells as opposed to the control.

Epinephrine is cytotoxic for both U87 and AGS cancer cells

Fig. 4 illustrates the cytotoxicity of the drug treatments after 24 h incubation with the drug agents. The data are expressed in percentage terms. Overall, both epinephrine and epinephrine/propranolol combinations were found to be cytotoxic, except for 16 µM epinephrine which interestingly was found to be non-toxic (Fig. 4A, for U87 cells) or even cancer cell-protective (Fig. 4B, for AGS cells). As with the outcomes of the previous experiments, epinephrine exhibited a dose-dependent activity reaching 40% and 25% cytotoxicity, for U87 and AGS cells (P<0.001). Also, AGS cells were found to be a little more resistant than U78 cells to this type of epinephrine activity. Concerning the combination therapy, propranolol displayed varied activity, from cytotoxic activity at the 16 µM epinephrine low concentration to the opposite cell-protecting activity at the highest 256 µM epinephrine concentration. In other words, the cytotoxicity of epinephrine at the 16 μM low concentration was significantly intensified in the presence of 75μM propranolol, it was augmented at 64 μM concentration mildly by propranolol, and it was gradually decreased at the concentrations higher than 128 μM by propranolol.

The present study investigated physiological and pharmacological epinephrine concentrations effects on the Both AGS (Stomach Cancer Cells-SCCs) and U87 (Brain Glioblastoma Cancer Cells- BGCCs) cell lines which included cell proliferation, adhesion, and viability in two levels of proliferation immittances (High-speed proliferation rate for AGS cell line, survival rate less than one year, and Low-speed proliferation rate for U87 cell lines, survival rate more than years). The results have shown that epinephrine at physiological concentration enhanced the proliferation of both AGS and U87 cells. However, pharmacological concentrations potentially decreased cell proliferation. High concentrations of epinephrine have shown toxic effects, which inhibit the proliferation of both cell lines in-vitro and may reduce tumor size in-vivo. It appears that increasing epinephrine concentration to more than 64µmol/L, oxidative stress leads to create hydrogen peroxide and reactive oxygen species (ROS), which provide cytotoxicity epinephrine characteristics. The results of this study are consistent with Behonick GS et al. and Costa V. M et al. who reported epinephrine may have toxic effects at doses above physiological levels (20).

These blocking effects of propranolol through β adrenergic receptors which are located on two cell lines membranes, could provide a reverse reaction to epinephrine in both low and high concentrations of agonists. Dong J and his colleagues reported that, after adding norepinephrine, in a concentration-dependent manner to glioma LN229 and U251 cells, due to the expression of both beta-1 and β2-adrenoceptors, proliferation was significantly enhanced and blocked by propranolol as a nonspecific beta-adrenergic receptor blocker functions (2).

The effect of epinephrine on the proliferation of HT-29 adenocarcinoma cells is create through both β1 and β2 adrenergic receptors demonstrated by Wong HP (20). Jun-fang Qin also reported that epinephrine promotes cancer through the β2 receptor by affecting the Tumor Micro Environment (TME) (21).

Oral squamous carcinoma facing increased levels of epinephrine (10µg/Ml,100µg/Ml) delayed scratch closure among cancer cells through inhibition of intracellular cAMP reported by Yamanaka (23). Also, Wounds healing assay by Raja Sivamani had shown that wounds under stress conditions, increased levels of epinephrine and expression of beta-adrenergic receptors on keratinocytes, resulting in impaired cellular epithelialization and delayed wound healing. On the other hand, treated beta-adrenergic antagonists (timolol) significantly increases the epithelial level of the wounds (24). Kiecolt-Glaser demonstrated that wounds of those who were most under psychological stress improved later than those without psychological problems (25). Loft and Klaunig et al. have shown oxidative DNAs play an important role in mutagenesis and increased risk of tumors (26). Djelic in cytogenetic experiments on human lymphocytes applied six experimental concentrations of adrenaline (0.01–200µmol/L) and reported that lower concentrations of epinephrine had no genotoxic effect on sister chromatid exchange and micronucleus. However, higher concentrations (5µmol/L,50µmol/L, 150µmol/L and 200µmol/L) due to the production of reactive oxygen species (ROS), decrease the mitotic index and delay the cell cycle (27).

Liu Y showed that epinephrine (5–50µmol/L) increased myeloma U266 cell growth in a dose-dependent manner after 24hrs and cell proliferation was inhibited by the β receptor antagonist, propranolol at a concentration (50–200µmol/L). The ratio of early and late apoptotic cells increased after treatment with propranolol. Caspase expression was increased in myeloma cells treated with propranolol (28). ÖD sahin FI et al. showed that propranolol at a concentration (of 100µmol/L) acts on tumor cells (breast MCF7, colon HT-29) by inducing apoptosis of cancer cell lines and stimulating the proteolytic activity of caspase-3 and caspase-9 significantly reduced cell migration, wound healing and declined proliferation (29).

Jóźwiak et al. have shown on human glioma C6 cells and human glioma T98G cells, adrenaline, and noradrenaline promote tumor growth by β1-adrenoceptors and cAMP activation. Rosenberg revealed that all tested catecholamines, including norepinephrine, epinephrine, and dopamine, were toxic on neurons as well as glia at 25µmol/L concentration. Oxidative degradation of catecholamines and hydrogen peroxide production, and adrenochrome (endogenous catecholamines) may play a role in normal and abnormal cell death. The toxicity of norepinephrine was blocked by catalase (30).

The present study also illustrated that in presence of propranolol the adhesion of U87 cells increased and metastasis decreased which means the epinephrine at physiological concentrations increases tumor growth while simultaneously preventing its metastasis. However, its pharmacological concentrations display toxic effects that decrease U87 cell proliferation and further enhance the metastatic state of tumors.

Numerous studies confirm these results. Palm, D showed that the migration of breast, prostate, and colon cancer cells was enhanced by stress-related neurotransmitters, norepinephrine (NE = 10µmol/L) in vitro, and this effect was inhibited by the inhibitor, β-propranolol (10µmol/L) (31). Barbieri A and colleagues showed that in the presence (NE = 10µmol/L), in vitro migratory activity of DU145 cells in prostate cancer increased and tumor cells from the primary tumor to the inguinal lymph nodes, resulting in the formation of larger metastases. Attention is strengthened on propranolol reduces metastasis by blocking norepinephrine function (32).

Pu J and colleagues reported that in a study of the PANC-1 pancreatic cancer cell model, epinephrine promotes migration in a dose-dependent manner and contributes to stress-induced metastasis in PANC-1 cells. By blocking β-adrenoceptor β2, cell migration was significantly reduced (33). Weng Mei. reported the effects of epinephrine on proliferation, migration, and invasion of glioblastoma U87MG and U251 cell lines with different concentrations of epinephrine (0.1,1,10,50,100µmol/L) after epinephrine treatment. The proliferation occurred only at U251. However, migration and invasion were significantly increased in both U87MG and U251 cells. Propranolol, (β-AR blocker) could reverse the effect of epinephrine on both cell lines. They also suggested that stimulation of human glioblastoma cell proliferation and invasion may be related to MMP-9 expression (13).

The present study also revealed that propranolol reduced cell viability at low concentrations of epinephrine and decreased the toxic effect of epinephrine at higher concentrations. The data show that at physiological concentrations of epinephrine, although increased the viability of U87 cells. However, pharmacological concentrations decrease cell viability and create toxicity effects of epinephrine. Other studies confirm our findings. Bustamante P et al. The antitumor effects of propranolol induce cell apoptosis in melanoma (UM) and cutaneous melanoma (CM), thereby reducing cell proliferation and migration, reduces the viability of cancer cells (34). Yoshioka Y et al. Showed that noradrenaline (10µmol/L) induces glutamate cysteine protein and increased glutathione (GSH) concentration by the protective role of astrocytes on human β-251 malignant glioma cells through adrenergic stimulation, increasing survival. This effect was inhibited by a non-selective β-adrenoceptor antagonist propranolol (35). According to Patri M and colleagues' results, in both brain tumor cell lines, such as neuroblastoma and glioma C6, norepinephrine increased cell viability by restoring the G2 phase of the cell cycle and decreasing the percentage of cell death (36). Zhou DR et al. reported the toxic effects of epinephrine due to stress induced by extracellular chemicals entering cells and disrupting cellular homeostasis and activating mitochondrial signaling cascades, resulting in increased steady-state levels of reactive oxygen species (ROS) and activation of Bax and caspases and cellular damage. This can ultimately lead to cell death and reduced survival (37).

Uchida et al. Showed that catecholamines, such as epinephrine above concentrations (60µmol/L), decreased the number of living cells in the Human Oral Squamous Cell Carcinoma lines due to ROS production and cytotoxicity cell. However, Catalases also reduced the toxicity of this effect of adrenergic agonists (38).

As the data of the present study indicate the dual effects of epinephrine due to the potential effects of its products, like ROS production is a long-standing issue in cancer, its toxic threshold can be an effective strategy to reduce tumor cell viability. On the other hand, many chemotherapy treatments kill the cells by increasing ROS concentration in the cell (39). Another study by Ciccarese F. showed that reactive oxygen species (ROS) act as a double-edged sword in cancer cells. Increased mtROS production leads to increased mitogenic signals, oncogenic transformation, genomic instability, and evasion of cell cycle inspections. On the other hand, excessive accumulation of H2O2 leads to irreversible protein modification, oxidative damage to lipids and nucleic acids, blockade of proliferative signaling, and ultimately cell death. Therefore, elevated ROS levels may reflect the Achilles' heel of cancer cells that can be therapeutically abused because it may overcome the toxic threshold by a slight increase in ROS levels, leading to mitochondrial crest regeneration and apoptotic cell death. High levels of ROS in cancer cells balance with increased antioxidant defense (40).

Taken together, the current study revealed that epinephrine serves anticancer pharmacological action at extremely high concentrations, due to the cytotoxic activity of the hormone at such aberrantly high concentrations. However, epinephrine is proven as a stress-related cancer-promoting hormone at low physiological concentrations, and propranolol as an antagonist beta-blocker reduces the cell viability, cell adhesion capability, and proliferation/migration rate at these concentrations. Epinephrine acts as a double-edged sword depending on the concentration; its low concentrations (physiological) increase cancer cell proliferation, enhance cell viability, and boost tumor growth, and its high concentrations (pharmacological) act vice versa. The addition of propranolol as a beta blocker reduced the cancer-promoting and invasive characteristics studied for epinephrine, especially at physiological concentrations for both slowly-growing U87 and fast-growing AGS cells.

The fabulous gain of this matter is that using a combination of epinephrine with other medicines might contribute to improved chemotherapy and cancer treatment. Propanolol is shown to hamper the cancer-promoting features of epinephrine at physiological concentrations. This indicates that sequential treatment of cancer patients with epinephrine and propranolol might lead to cancer/tumor treatment, especially when the epinephrine dose is reduced in the body. Combination therapy of epinephrine with chemoagents at the right stage of tumor development may have higher effects on destroying cancer cells, and lead to a patient’s recovery and healthy conditions with fewer sessions of chemotherapy. This means a shorter period of treatment in the NHS system by the government and private sector and more hope for healing patients even with higher grades of cancer tumors.

Conflict of Interest

The authors declare no conflict of interest.

Author Contribution

M.M., N.R, A.N. and A.M.S. collected the data, M.M., J.A. , M.T and S.A.K. performed the statistical analyses, interpreted data, and drafted and revised the manuscript for important intellectual content.H.Z., N.R. and A.M.S. reviewed the analyses and the final version of the manuscript. M.M., M.T and A.M.S. interpreted data, revised the manuscript for important intellectual content, and approved the final version.All authors have read and approved the manuscript.

Acknowledgement

This study was a part of Abbas Noori’s M.Sc. dissertation and financially supported by grant number 1120, with the ethical code IR. MAZUMS.REC.1397.231 from Immunogenetics Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran

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No competing interests reported.

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The Dual Influence of Adrenaline may be Effective in the Treatment of Grade 3 and 4 Cancers with a Synergistic Effect with Chemotherapy

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Abstract

Figures

Introduction

Material and methods

Materials

Cell growing conditions

Treatment groups

Scratch wound healing assay

Cell adhesion assay

Cell viability

Statistical analysis

Results

Epinephrine reduces the cells’ proliferation/migration rate

Epinephrine increases the number of detached cells

Epinephrine is cytotoxic for both U87 and AGS cancer cells

Discussion

Conclusion

Declarations

Conflict of Interest

Author Contribution

Acknowledgement

References

Additional Declarations

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