Artemisia: A Promising Herbal Approach for The Treatment of Cancer

Harshadeep Seal*, Namrata Koushik

Department of pharmaceutical science, Girijananda Chowdhury University, Dekargaon, Tezpur, Assam, 784501

*Correspondence: harshadeepseal150132@gmail.com;

INTRODUCTION

Artemisia is an herbal drug which, has been long used in Chinese traditions, scientists found newly use in treatment of various diseases, malaria and cancer being the most popular among them. It’s a plant which has many species currently in study, mainly Artemisia annua and Artemisia vulgaris. Artemisia is a plant of asteraceae family. It is indigenous to Asia however, localized in many other countries including North America, Europe etc. Based on Chinese traditions, indigenous people has treated people with various disease and is said to be effective against fever, acanthamoebiasis, cancer, schistosomiasis, HIV, hepatitis-B, Leishmaniasis, etc. (2). Recent study illustrated that this plant possesses high levels of phenolic compounds like sesquiterpene lactone (artemisinin), coumarin, flavones, flavonols, flavonoids and phenolic acids. Since, researches have been carried out regarding effect of artemisinin and flavonoids with cancer, the main objective of this study is to review the anti-cancer properties of this plant (2, 3, and 4).

Objective of the study

This overview is to review the chemical compounds of Artemisia annua and Artemisia vulgaris as well as the anti-cancer properties in details.

MATERIAL AND METHODS

This systematic review was performed by searching for studies in the databases PubMed and Google scholar. The initial search strategy identified approximately 20 references. In this study, 4 studies were accepted for further reviews and met all of our criteria.

All the parts of each article which consisted of the title, abstract, introduction, methods, results, discussion, and conclusion were looked upon and studied.

RESULTS AND DISCUSSION

Inhibitory effect of artemisinin with pKAL against advanced metastatic breast cancer cell

 The effect of artemisinin combined with pKAL (a type of polyphenol) on advanced metastatic breast cancer cell was studied. Here, pKAL inhibited cancer metastatis, which is secondary malignant growth. Its mechanism involved inhibiting cancer cell adhesion to ECs (endothelial cells). Extended term treatment by Artemisia and bicalitumide (type of anti-androgen which block effects of testosterone, which fuels the growth of prostate cancer cells) showed inhibition of metastasized prostate carcinoma (1).

Anti-tumor activity of Artemisia annua against human cancer cells

 Effect of Artemisia were also seen on three human cancer cells, from which F7 (purine derivative drug used for enhancing targeting efficiency) combined with Artemisia displayed potent action against SK-N-MC cells (which is a kind of human neuroblastoma cell line). F7 with DOX (doxorubicin) increased the anti-tumor activity and cytotoxicity of DOX by regulating activity of multi-drug resistant cancer cells through apoptosis (1).

Factors influencing anti-proliferative activity of Artemisia

 The anti-proliferative action of the plant’s extract was studied and it was seen that the anti-tumor activity of Artemisia not only depends on varying concentration of active ingredients present in its oil but also depends on the site of treatment ( or target site ) and different parts of the plants used to extract the active ingredient (1).

Anti-cancer effect of arsantin in human promyelocytic leukemia cell

Arsantin (an active ingredient of Artemisia annua) displayed activity against human promyelocytic cells (HPL cells). Arsantin effectively increased difference between HL-60 cells and its effectiveness increased with respect to the dosage concentration (1).

Inhibitory effects of Artemisia annua on osteosarcoma:-

Study on osteosarcoma was also done, where Artemisia affected the cell’s proliferation, apoptosis factor and metastasis (1).

 Anti-proliferative action of Artemisia against melanoma cells

 An in-vitro test was done, in which effect of Artemisia was seen on melanoma cells. Results showed the proliferative inhibition of melanoma cells in concentration related manner and it increased over expression of both BAX and cytochrome C. The BAX activation induced mitochondrial membrane permeabilization, thereby leading to the release of apoptotic factor cytochrome C which inhibited cancer cell proliferation (1).

High taxadiene producing genetically engineered plant of Artemisia annua

 Genetically engineered plants of Artemisia were also studied. These biologically engineered plants produce more taxadiene than the normal ones. Taxadiene is an essential precursor for the biosynthesis of highly potent anticancer drug paclitaxel. These transgenic plants were very useful since the general production of taxadiene has a very low yield (which is done with the help of microbes). The outcome showed the possibility of biological engineering of the biosynthetic taxane pathway in Artemisia annua (1).

Anti-austerity activities of Artemisia vulgaris against PANC-1 human pancreatic cancer cell line

Preferential toxicity was shown against the PANC-1 human pancreatic cells by 70% ethanolic extract of Artemisia vulgaris in NDM (nutrient deprived medium). The PC50 (the concentration at which 50% of the pancreatic cancer cell was killed in NDM without showing toxicity in DMEM medium) was found out to be 12.5 µg/ml (2).

 After phytochemical screening, a total of 9 compounds were found present in the Artemisia extract. Among them, a new sesquiterpene compound was seen and it was vulgaric acid. All these reported compounds were isolated and were tested for their preferential cytotoxic activity against the PANC-1 human pancreatic cells in NDM and DMEM. The results showed moderate preferential cytotoxic activity for all the compounds except apigenin. Apigenin had the strongest PC50 value of 30.7 µg/ml (2).

Efficacy of dried Artemisia annua leaf extract against Non-small Cell Lung Cancer (NSCLC)

The effect of AS (artesunate) and DLA (dried leaf Artemisia) extract on cancer cells were compared. Generally, cancer cells have higher percentage of iron compared to normal human cells since, their iron intake is higher than the normal. The sesquiterpene lactone present in Artemisia like artemisinin, artesunate, artemether etc. has an endoperoxide bridge present in their structure which react with the free ferrous iron and generate reactive oxygen species (ROS) which are highly damaging to the cells. Higher iron content in cancer cells makes them more vulnerable to damage by ROS compared to healthy cells (3).

a) Cytotoxic effects of AS and DLA extract on NSCLC cell lines:

Treatment of NSCLC cells with AS and DLA extract showed apoptosis in the cells. MTT assay was done which displayed dosage concentration-dependent inhibition of proliferation in the cell lines for both AS and DLA extract. The IC50 were found out to be in µM (micromolar) range for both. They did not show any significant effect on the solvent used (either DMSO or MeCl₂). Histone H2AX protein phosphorylation increased in all the NSCLC cell lines when treated with either AS or DLA extract. It indicated that the cell death could have occurred by crossing the threshold of repairable DNA damage was crossed (3).

b)  Cell cycle arrest in NSCLC cells:-

Cell numbers increased in G₂/M phase when AS or DLA extract were employed in PC9 and H1299 cells (types of NSCLC cell line). PC9 cells increased by 46.4% when treated with 50 µM AS whereas, it increased by 54% for 50 µM DLA extract. AS at 100 µM (value of concentration close to IC50 value) increased cell numbers in G₂/M by 53.3%. H1299 cells after being exposed to AS and DLA extract at 50 µM concentration increased the cell numbers by 36.5% and 46.7% respectively in G₂/M phase. In addition, at 75 µM it increased the number of H1299 cells by 39.5% and 47.2% for AS and DLA extract respectively. Treatment with 150 µM AS increased A549 cells (type of NSCLC cell line) in G₁ phase by 56.2% which was the same as control whereas, 150 µM DLA increased it by 68.4% (3).

Hence, both of them AS and DLA extract caused G₂/M phase cell cycle arrest in PC9 and H1299 cells. However, only DLA extract induced G₁ phase cell cycle arrest in A549 cells. To confirm this outcome, phosphorylation of histone H₃ at Ser 10 in western blot was done. It displayed increase in phospho-H₃-Ser 10 protein in AS and DLA extract treated PC9 and H1299 cells (3).

c) Effect of migratory ability in NSCLC cells by AS and DLA extract

To check the effect of change in migratory ability, wound-healing assay showed that DLA extract reduced the number of migratory PC9 and A549 cells into the wound by half (50%). However, it did not restrict the migration of H1299 cells (3).

d) Caspase -3, -8 and -9 activation in NSCLC cells when exposed to AS and DLA extract:-

The cleavage of caspase occurs during apoptotic pathway. AS and DLA extract induced caspase activation and cleavage. Here, cleaved caspase -3.-8 and -9 were studied for involvement of intrinsic and extrinsic apoptotic pathway. AS and DLA extract approximately in its IC50 increased response of protein levels in cleaved caspase -9 and -3 compared to control in all NSCLC cells. Protein levels of cleaved caspase -8 also increased in A549 and PC9 cells, but not in H1299 cells (3).

e) DLA extract suppressed tumor growth in NSCLC murine xenografts

After p.o (per os) delivery of AS or DLA extract in vivo, inhibition of tumor growth took place significantly on day 14 in A549 xenograft models. Moreover, it reduced the overall volume of the tumor. However, DLA extract treated xenograft had 50% reduction in its tumor growth which when compared to AS treated xenograft wasn’t that significant (3).

During this 14-day treatment, the animal weight remained stable throughout (3).

Identification of active ingredients of an Artemisia annua herbal preparation and its anti-tumor activity

Generally, the sesquiterpene lactone artemisinin is known to be the active ingredient of Artemisia. However, it was showed that artemisinin might not be the best anti-cancer compound present in Artemisia annua. Other compounds present in the plant can be highly effective against cancer cells. These compounds displayed potent activity against highly metastatic triple-negative breast cancer (TNBC) cells (4).

Here, Momundo extract and acetonitrile (ACN) macerated Momundo extract were choosen. Momundo Artemisia annua extract (PZN 5466281) was obtained from MoMundo GmbH (Bad Emstal, Germany). These extracts were characterized by HPLC and they does not contain artemisinin in them. The major constituent present were 6,7- Dimethyl- coumarin, chrysosphenol, castacin, arteannuin B and arteannunic acid (4).

During the morphological analysis by fluorescence microscopy after 24 hour of treatment, it was found that Momundo-ACN extract induced formation of multinucleated cells, mostly 4N cells similar to potent anti-cancer drug paclitaxel. During this period, Momundo extracts did not show any kind of toxicity towards the cancer cells however, a slight increase in the apoptotic cell number was seen. To confirm the activity, cell cycle analysis was performed. It confirmed that, treatment with Artemisia extracts increased the number of tetraploid (4N) MDA-MB-231 (triple-negative breast cancer) cells and of those in S-phase of the cell cycle (4).

Momundo Artemisia extracts showed inhibition of MDA-MB-231 cells in a concentration and time dependent manner. Momundo-ACN extracts had comparatively higher cytotoxicity towards MDA-MB-231 cells. IC50 of Momundo-ACN and Momundo extracts were found out to be 18 µg/ml and greater than 100 µg/ml respectively after 48 hours of exposure (4).

To determine whether the cytotoxicity of Momundo Artemisia annua is due to apoptosis induction, various apoptotic parameters were analysed. Indeed it was, as number of cells with hypodiploid DNA contents treated for 48 hours with Momundo-ACN extract were significantly increased (which is similar to the mechanism followed by drug paclitaxel). Mitochondria further increased apoptosis by releasing pro-apoptotic proteins into the cytoplasm activating caspase -3. To demonstrate involvement of the intrinsic mitochondrial pathway in Artemisia annua induced apoptosis, analysis of the mitochondrial membrane potential was done. When Momundo extracts were used against breast cancer cells, it increased the percentage of cell bond, reduced membrane potential 4.0 to 18.1 % and by 43.4% respectively (similar to drug paclitaxel). It verified microbial membrane permeabilization. Also, Momundo extracts induced caspase -3 activation which confirmed induction of apoptosis. However, activation of the extrinsic pathway could not be inhibited (4).

MDA-MB-231 breast cancer xenografts were used to confirm the anti-tumor activity of Momundo extracts (invivo). Treatment of the xenografts with Momundo extracts dose-dependently reduced the tumor volume. Immunological analysis of tumors revealed the inhibitory action of Artemisia annua extracts against cancer cell proliferation. Therefore, treatment with either of the Momundo extracts reduced significantly expression of proliferation marker Ki-67. Also, these extracts increased the number of apoptotic TUNEL-positive cells (Terminal deoxynucleotidyl transferase dUTP nick-end labelling), which is a kind of assay indicating induction of DNA strand breakage and apoptosis in-vivo. However, no obvious systemic toxicity on the chicken embryo was observed (4).

Anti-cancer activity of Momundo extracts were also analysed in an orthotopic breast cancer model in mice. Daily treatment of animals with Momundo extract for three weeks retarded tumor growth of breast cancer xenografts (similar to standard chemotherapeutic drug doxorubicin). However, because of high toxicity of doxorubicin compared to Momundo extracts, doxorubicin was administered once weekly (4).

There was no effect on cancer cells due to arteannunic acid and 6,7- Dimethoxycoumarin. Arteannuin B, casticin, and chrysosphenol D strongly inhibited MDA-MB-231 cell viability. This indicates, although Momundo Artemisia extracts contain no artemisinin, it contains other natural compounds with potent cytotoxic and inhibitory activity against TNBC cells (4).

CONCLUSION

In this study, anti-cancer properties of Artemisia were overviewed. Findings of this study showed that artemisinin, a sesquiterpene lactone found in this plant can be effective against various cancer cell lines. Other compounds in this plant such as apigenin, arteannuin B etc. can also be if not more, equally efficacious against cancer. However, more studies are required to find new compounds involving similar properties and the side-effects of the compounds found in the plant if any.

ACKNOWLEDGEMENT

We appreciate the Department of Pharmacognosy of Girijananda Chowdhury University, Tezpur, Assam, for their support. In addition, we thank all those who cooperated with us in fulfilling this study.

REFERENCES

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