HeLa Cell Line

A member of genus Polyalthia, which has not been studied extensively is Polyalthia glauca. This study investigated chloroform extract from leaves of P. glauca with anticancer activities on human cervical adenocarcinoma cell line (HeLa) in vitro. The chloroform extract of P. glauca leaf was obtained by maceration. Further, the extract was tested using MTT assay against HeLa cell line. The cell cycle analysis was later conducted using flow cytometry, while the measurement of apoptosis was carried out through DNA fragmentation in HeLa cell treatment. The cytotoxicity test results showed that the IC 50 values of chloroform extract of P. glauca leaf against HeLa cells was

8. Anti cancer activity (MTT assay): The anticancer activity of ferulic acid was studied against HeLa cell line. HeLa cell line was obtained from National Centre for Cell Science (NCCS), Pune, India. The cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM) with 10% Fetal Bovine Serum (FBS), penicillin (100 U/ml), and streptomycin (100 μg/ml) in a humidified atmosphere of 50 μg/ml CO 2 at 37 °C. The cells were grown in 24-well plates

The Hela cell line was purchased from the cell bank of Pasteur Institute of Iran, and was placed in a culture medium containing RPMI 1640 supplem- ented with FBS 10% and penicillin/streptomycin 1% in an incubator under normal circumstances (37 °C temperature, containing 5% carbon dioxide, 95% moisture). Human dermal fibroblasts were enzyme- aticcally extracted from foreskin of 1-1.5 month-old infants, and were placed in a culture medium containing RPMI 1640 supplemented with FBS 10% and penicillin/ streptomycin 1%, in a CO 2 incubator

ABSTRACT: In present study antiproliferative activity of Rutin was evaluated on HeLa cell line induced cervical cancer in rats. For this study, 30 rats were divided into 5 groups and each group containing 6 rats each. Group I- normal saline treatment for 45 days, Group II- cancer cells (1×10 6 cells in 0.1ml/rat), Group III– 5- Fluorouracil (20mg/kg + 1×10 6 cells in 0.1ml/rat), Group IV- Rutin (50mg/kg + 1×10 6 cells in 0.1ml/rat), Group V- Rutin (70mg/kg + 1×10 6 cells in 0.1ml/rat). After 24 h of tumour inoculation intraperitoneally, Rutin was administered daily for 45 days. After administration of last dose followed by 18 hrs fasting, rats were sacrificed for observation of antiproliferative activity. The change in body weight, body circumference of tumour bearing hosts and simultaneous alterations in haematological profile, serum (Triglycerides, Total protein, Total cholesterol, GGT, ALP and glucose) and liver biochemical parameters (lipid peroxidation, GSH and antioxidant enzymes-CAT, GPx) were estimated. The changes in tissue enzymes- Glucose-6 phosphate dehydrogenase, Hexokinase, Succinate dehydrogenase and CytochromeP450 levels were also estimated. Rutin maintained the body circumference and body weight of proliferation bearing rat. Haematological profile reverted towards normal levels in Rutin treated rat. Treatment with Rutin restored serum biochemical parameters towards normal levels and decreased levels of lipid peroxidation and increased levels of reduced glutathione and other antioxidant enzymes. The Rutin treatment restored Glucose-6 phosphate dehydrogenase, Hexokinase, Succinate dehydrogenase and CytochromeP450 levels in proliferation induced rat. Rutin exhibited antiproliferative effect by modulating haematological parameters, lipid peroxidation and augmenting antioxidant defense system in proliferation bearing rat.

Background: Cervical cancer is the second most common female cancer worldwide. Inhibitors of apoptosis proteins (IAPs) block apoptosis; therefore, therapeutic strategies targeting IAPs have attracted the interest of researchers in recent years. Apollon, a member of IAPs, inhibits apoptosis and cell death. RNA interference is a pathway in which small interfering RNA (siRNA) or shRNA (short hairpin RNA) inactivates the expression of target genes. The purpose of this study was to determine the effect of constructed shRNAs on apoptosis and growth inhibition through the suppression of apollon mRNA in HeLa cell line. Methods: Three shRNAs with binding ability to three different target sites of the first region of apollon gene were designed and cloned in pRNAin-H1.2/Neo vector. shRNA plasmids were then transfected in HeLa cells using electroporation. Down-regulation effects of apollon and the viability of HeLa cells were analyzed by RT-PCR, lactate dehydrogenase assay, and MTT assay, respectively. Also, the induction and morphological markers of apoptosis were evaluated by caspase assay and immunocytochemistry method. Results: The expression of shRNA in HeLa cells caused a significant decrease in the level of apollon mRNA1. In addition, shRNA1 effectively increased the mRNA level of Smac (as the antagonist of apollon), reduced the viability of HeLa cells and exhibited immunocytochemical apoptotic markers in this cell line. Conclusion: Apollon gene silencing can induce apoptosis and growth impairment in HeLa cells. In this regard, apollon can be considered a candidate therapeutic target in HeLa cells as a positive human papillomavirus cancer cell line. DOI: 10.7508/ibj.2016.03.003

The objective of the study was to analyze the anticancer property of the leaves of Triticum aestivum on HeLa cells. The Indian medicinal plant Triticum aestivum that is used in traditional medicine for cancer and non cancerous diseases was collected. The crude aqueous extract was prepared by using standard protocols. The antiproliferative effect of the aqueous extract was evaluated in vitro by employing MTT assay. The potency of each plant extract concentration was calculated in terms of percent cell inhibition of VERO and HeLa cells. The extract showed dose dependent anticancer activity on the cancer cell line i.e HeLa cell line while the extract did not show any cell toxic potential to the normal cell line i.e. Vero cell line. . The MTT assay showed an anti proliferative activity (IC 50 ) for the HeLa cell line at 133.6 μg/ml of crude extract.

Research in the search for cancer drug compounds continues to be developed, considering that specific anticancer compound has not been obtained. Some of the urea derivative compounds are also being developed in the search for an anti-cancer compound which is potent with minimal side effects. Related to the explanation above, a urea derivative, which is N-phenylurea, would like to be developed. It would be reacted with 4-methoxybenzoyl chloride, thus N-4- methoxybenzoyl-N'-phenylurea compound would be obtained. The synthesis of N-4-methoxybenzoyl-N'-phenylurea was carried out by Schotten-Baumman method that had been modified, and then a test of purity was done by thin layer chromatography using 3 different kinds of eluent. The next phase was structure characterization which was carried out by using UV and IR spectrophotometry, then 1 H-NMR spectrometry also MS, so that the structure from N-4- methoxybenzoyl-N'-phenylurea would be obtained. Anticancer activation test was conducted on HeLa cell line by using MTT assay, and the value of IC 50 would be obtained. The

The antibacterial activities of the microbial pigment of the actinomycetes were carried out against the bacteria such as Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Escherichia coli, Salmonella typhi. The E. coli (8 mm) more susceptible than other microbial pathogens. Whereas remaining species showed negative result. The antifungal activity of the microbial pigment of the actinomycetes were carried out against the four different fungi such as Candida albicans, Aspergillus niger, Trichoderma viride, Rhizopus microporous. The actinomycetes pigment were failed to inhibit the pathogenic fungi. There is no antifungal activity against the all tested pathogenic fungi. Fourier Transform Infra-Red (FTIR) spectrum revealed the NH and NOH stretching for the actinomycetes pigment. In vitro cytotoxic assay on Human Cervical Adenocarcinoma (HeLa) cell line. The actinomycetes pigment prodigiosin, tripyrrole ring, was found to exhibit antiproliferative property against the (HeLa cell line. The pigment was found to be effective against the HeLa cell line. The IC 50 value of the pigment was 62.50

Andrographiselongata (Vahl) T. Anderson or Mullurinjipachilla is an endemic medicinal plant. The plant is usually used against many ailments in old Travancore. The plant part is used in the treatment of snake bite, diabetes, cough, skin diseases. The main objective of this study is to identify the scientific basis of this plant used in traditional practices. In this study we analyze the anti- proliferative effect of the leaves of A. elongata on HeLa cell line. The cytotoxicity of the methanol leaf extract was evaluated in vitro by employing MTT assay. The extract showed dose dependent anti-proliferative activity with an IC 50 of <100 µg/ml in HeLa cell line.

Cell Culture: HeLa (Human Cervix Adeno carcinoma), human lung carcinoma cell line (SK- LU-1), human colon carcinoma cell line (HCT-116), and Fibroblastwere used. HeLa cell line was cultured in Minimum Essential Medium (MEM) with supplemented with 5% of FBS. SK-LU-1 and HCT-116 cell line were cultured in Eagle's minimal essential medium (EMEM) supplemented with 10% of FBS. Fibroblast cell line was cultured in Dulbecco's modification of Eagle's medium (DMEM) supplemented with 10% of FBS. T-25 flask was placed in laminar flow and the spent cultured medium was discarded with a sterile Pasteur pipette. A total volume of 2mL of 1xPBS was added to the side of the T-25 flask opposite the cells to avoid dislodging the cells. Then, the cells were rinsed and the rinse was discarded afterwards. A total volume of 2mL of trypsin solution was added to the cells, ensuring the monolayer is completely covered. The T-25 flask was incubated for 5-10 minutes or the flask gently tilted until the monolayer can be seen detaching from the culture surface. Cells then observed under microscope and resulting cells should be rounded and floating. A total volume of 2mL of medium and cells were dispersed gently by repeated pipetting. Subculture was performed in a ratio of 1:4 (cell suspension:

In medical research, the most famous immortal cell line known as HeLa was developed from cervical cancer cells of a woman named Henrietta Lacks.HeLa cell line was obtained from National Centre for Cell Science(NCCS) Pune and maintained in Dulbecco’s modified Eagle’s media (HIMEDIA) supplemented with 10% FBS (Fetal Bovine Serum) and grown to confluence at 37°c in 5 % CO 2 in a humidified atmosphere in a CO 2 incubator(NBS, EPPENDORF, GERMANY). The

few sequence features that enable robust computational prediction of bacterial effector proteins. This has led to a need to screen for proteins via their effects upon model hosts. Initially such screens were performed using libraries of bacterial mutants 8,9 , but this often fails to find all secreted proteins due to trans-complementation between mutants within a pool, or effector redundancy. More direct approaches such as expression of putative effector proteins in yeast have been able to find new effectors, place their effects within host pathways 10,11 , and more recently demonstrate the synergy and interactions between effectors 12 . In some cases these observations have been extended to other infection models or mammalian cell systems 13 . However, in yeast the primary readout is growth – whilst this makes screening robust and cheap, it gives little additional information on which cellular pathways are involved in effector action, although recently this has been improved with subcellular localisation screening of known secreted effectors in yeast 14 . Extending this approach to mammalian cells, we are able to gain more information by using reporters in pathways of interest, as well as simultaneously imaging a number of cellular components and cell properties. Using fluorescently-tagged bacterial proteins also shows the localisation of specific bacterial proteins within the host cells, offering further insight into their roles and functions. A similar approach has been recently used for assaying human gene function and has been termed cell painting 15 .

Infection with most cytolytic animal viruses is characterized by a marked inhibition of host transcription and translation (5, 58). This inhibition is particularly evident when HeLa cells are infected with poliovirus (9, 16). The selective blockade of mac- romolecular synthesis in virus-infected cells is accompanied by massive poliovirus RNA replication and almost exclusive syn- thesis of viral proteins. Poliovirus RNA translation starts at a single AUG initiation codon and continues through a long open reading frame that encodes a huge polyprotein precursor (67). This precursor is not found as such in the infected cells, since it is proteolytically processed while still attached to ribo- somes as a growing peptide (32, 51). Two virus-encoded pro- teases, 2A pro and 3C pro (or its precursor 3CD pro ), are respon-

Chemoprevention by dietary constituents has a valuable role in the control of diverse diseases including cancer (Kontou et al., 2011). Camel milk is an example for an excellent source of these constituents that exhibit different biological activities (Yagil et al., 1982). In the present study camel milk and its whey proteins emerged as a powerful anticancer agents that reduced the in vitro growth of Hela cells. According to the obtained results, they showed a toxic effect on the studied cells that approximately reached 85%. Apparently, camel milk is one of the natural products that has cytotoxic potential against cancer cells like murine hepatoma hepa 1c1c7 cells (Korashy et al., 2012). In the running study, the cytoxicity of camel milk and whey could be referred to the presence of casein, lactoferrin and immunoglobulins (Konuspayeva et al., 2007). It was recorded that camel milk increased the expression of chemo-protective genes which in turn increased the levels of several antioxidant enzymes. These enzymes prevent the formation of highly reactive oxygen species and so protect DNA and cell damage (Habib et al., 2013). Other study showed that casein and its product formed during pepsin hydrolysis had the ability to protect mammalian cells against certain genotoxic compounds (Van Boekel et al., 1993). Moreover, lactoferrin, the other component of camel milk exerts antitumor activity due to the immune- inducing and immunomodulatory properties (Al-Majali et al., 2007).

hexanucleotide repeats, which form the caps at the chromosome ends, is implicated to determine the aging process, and more importantly the healthy lifespan itself. Telomerase, a ribonucleo- protein having reverse transcriptase activity, arrests telomere loss through addition of the TTAGGG repeats de novo, to the ends of the chromosome. The telomere/telomerase maintenance is an inevitable necessity to delay aging and for a healthy lifespan. Here, we report the potential of full-spectrum, high concentration Ashwagandha (Withania somnifera), an Ayurvedic medicinal herb, root extract to increase telomerase activity. HeLa cells, when treated with various concen- trations of Ashwagandha root extract, showed an increase in telomerase activity measured with the established Telomerase Rapid Amplification Protocol (TRAP) assay. Ashwagandha root extract increased telomerase activity with highest enhancement of ~45% at 10 - 50 µg concentration. Thus, Ashwagandha root extract has the anti-aging inducing potential.

The docking simulation of 3,7,11-trimethyl-3-dodecanol with Prepared Heat Shock protein 70 had a glide score of -6.27. Upon the examination of docking features between 3,7,11- trimethyl-3-dodecanol with Prepared Heat Shock protein 70, one hydrogen bond was formed between the 3,7,11-trimethyl- 3-dodecanol with Prepared Heat Shock protein 70 with LYS 254 at 2.06A (Fig 8).Comparison among all the interactions observed, the binding simulation with Heat shock protein 70 as a cancer protein target shows a higher Glide score indicating the stability in interaction. The HSP70 family of proteins can be thought of as a potent buffering system for proteotoxic stress, cellular stress, either from extrinsic like physiological, viral and environmental or intrinsic like replicative stimuli. As such, this family very essential for survival functions in the cell. Remarkably, cancer cells absolutely rely on the buffering system for continued survival. (Elisa Zorzi et al., 2011) It has been acknowledged for many years that HSP70 is frequently over expressed in transformed or cancerous cells. Cancer cells over expressed few types of protein in cytoplasm, high levels of proteotoxic stress in tumors and subsequent activation of HSF1, which further contributes to the frequent over expression of the HSP70 gene in cancer cells. (Sandy et al., 2009)In habitually cytochrome c in mitochondria is liable for external and intrinsic stimuli for apoptosis. Mitochondria DNA is responsible for apoptosome release via cytochrome c. It releases the quaternary apoptosomeproteins to the cytoplasm, Apoptotic proteinase activating factor1 interconnected to apoptosome and then binds with deoxyadenosine triphosphate factor. Interacted entire factors can revert the inactivated pro caspase 9 to activate scaspase

Culturing of tissues in a favorable artificial environment is called tissue culture. Tissue culture is divided into two types: (i) primary cultures and (ii) secondary cultures. Primary cell culture is the maintenance of growth of cells dissociated from the parental tissue. The (I) Primary cell culture could be of two types depending upon the kind of cells in culture (Jacoby et al., 1979). They are Adherent cells and Suspension cells. Adherent Cells require attachment for growth are said to be anchorage dependent cells and Suspension Cells which do not require attachment for growth are anchorage independent cells/suspension cells. For example is lymphocyte. (ii) Secondary cell cultures: When a primary culture is subcultured, it is known as secondary culture or cell line. A cell line or cell strain may be finite or continuous depending upon whether it has limited culture life span or it is immortal in culture. On the basis of the life span of culture, the cell lines are categorized into two types. One is Finite cell lines and another one is infinite cell line. The Cell lines which have a limited life span and go through a limited number of cell generations (usually 20-80 population doublings) are known as finite cell lines. The Cell lines which are transformed under laboratory conditions or in vitro culture conditions give rise to continuous cell lines. They grow either in a monolayer or in suspension. The growth rate is rapid and doubling time can be 12-24 hours (Capes et al, 1993).

Introduction and aims Cervical cancer has become the third most common cancer among women and second most common cause of death. The current treatment modality has issues of drug resistance and side effects. Hence, investigation of plant species as a source of experimental therapeutic agents, in treating cancer is currently gaining a lot of importance. One such naturally available plant extract is Melaleuka alternifolia (TTO) which belongs to the family of essential oils is a very good antibacterial, antifungal, antiviral, antiprotozoal and anti-inflammatory agent. But currently there is a lot of importance is given for its anticancer effect. Hence our aim is to evaluate anticancer activity of Melaleuka alternifolia on cervical cancer cell line (HeLa) by MTT assay an in vitro method. Methodology: Before the start of the study ethical clearance was obtained from Institutional Review Board. The cytotoxicity checked for cervical cancer (Hela) cell line and Vero Monkey kidney cell line which was used as a control in our current study. These cell lines were procured from NCCS Pune, India. 1. MTT solution preparation (stock solution): 5 mg in 1 ml of PBS. 2. Cell culture : The cell lines were maintained in 96 wells micro titer plate containing MEM media supplemented with 10% heat inactivated fetal calf serum (FCS), containing 5% of mixture of Gentamicin (10ug), Penicillin ( 100 Units/ ml) and Streptomycin (100µg/ml) in presence of 5% CO2 at 37ºC for 48-72 hours. 3. Cytotoxicity Assay: In vitro growth inhibition effect of test compound was assessed by calorimetric or spectrophotometric determination of conversion of MTT into Formazan blue by living cells. Results The results represent the mean of five readings. The IC50 value of tea tree oil for cervical cancer cell line after 48 hrs was 3.125µg/ml. Spearmans rho’s Correlation showed P value <0.05 indicating there was statistical significant results obtained when TTO was treated with HeLa cervical cancer cell line for 48 hrs incubation period. Conclusion: TTO has a promising anticancer property against cervical cancer cell line (HeLa) with its IC50 value 3.125µg/ml. Hence this TTO with its greater efficacy related to its anticancer activity can be brought to the level of clinical trials in the coming future.

Few of the selected compounds were tested for their cytotoxic activity against human cervical cancer HeLa cell line and squamous cell carcinoma of human skin HSC-1 cell line. All the tested compounds were active against both the cell lines. The compounds containing flurophenyl 7, nitrophenyl 9 and chlorophenyl substituents 10 have shown good cytotoxic activity for both HeLa cell line and HSC-1 cell lines. It indicates that the activity of the tested compounds was influenced considerably by the nature of the substituents on the aryl group. Among the tested compounds, 7 can be identified as the most promising compound against both cancer cell lines.

This paper presents a prototype tool for generating textual definitions for an ontology from logical definitions using the Experimental Factor Ontology (EFO) [1] as a case study. The heart of ontology building is the definition of entities in a domain. A defini- tion states what kind of thing the described entity is and how it is distinguished from other entities of the same kind. As such, a definition states how an entity can be distinguished or recognised from other entities. Such definitions come in two styles with a common core aim: natural language or text definitions of an entity and logical definitions of an entity. Figure 1 shows an axiomatic description in OWL and a hand- written textual definition for the HeLa cell line from EFO (left and central panes). The information within the two types of definition is similar (they both talk of cells that come from a human cervical carcinoma; the hand-written, however, also gives the information of the individual human whence the cells came), but they differ in style of rendering and apparent ease of reading.