Viability Of HeLa Cells Research Articles

TLyP-1-conjugated core-shell nanoparticles, Fe3O4NPs@mSiO2, for tumor-targeted drug delivery

Tumor-targeted core-shell nanoparticles were developed for a multifunctional drug delivery system (DDS) for cancer therapy. Fe3O4 nanoparticles with mesoporous silica (Fe3O4NPs@mSiO2), in which Fe3O4 and mSiO2 form the core and shell, respectively, were functionalized to deliver a hydrophobic anti-tumor drug and to heat targeted tumor cells with the energy of an alternating magnetic field. For targeting tumor cells, tLyP-1, a tumor cell homing and penetrating peptide, was engrafted on to the Fe3O4NPs@mSiO2 (Fe3O4NPs@mSiO2-tLyP-1). The fabricated Fe3O4NPs@mSiO2-tLyP-1 were loaded with camptothecin (CPT); they showed robust, selective targeting and penetrating efficacy for Hela cells, and induced cell death. The CPT-loaded Fe3O4NPs@mSiO2-tLyP-1 (Fe3O4NPs@mSiO2-tLyP-1(CPT)) reduced the cell viability of Hela cells to 28.8%. Furthermore, in combination with application of an alternating magnetic field to cause hyperthermia, the nanoparticles made it possible to decrease viability to 18.3%. The systemic toxicity of Fe3O4NPs@mSiO2-tLyP-1(CPT) to human mesenchymal stem cells (hMSCs) was minimal, because the nanoparticles selectively targeted and penetrated the tumor cells. This study indicates that the developed Fe3O4NPs@mSiO2-tLyP-1 have potential as a DDS in the chemo-hyperthermic treatment of cancer.

Could FA-PG-SPIONs act as a hyperthermia sensitizing agent? An in vitro study

The therapeutic effect of polyglycerol coated iron oxide nanoparticles (PG-SPIONs, non-targeted nanoparticles) and folic acid-conjugated polyglycerol coated iron oxide nanoparticles (FA-PG-SPIONs, targeted nanoparticles) in combination with hyperthermia on viability of HeLa cells was investigated. It was observed that coated and uncoated SPIONs have spherical shapes with an average diameter of 17.9 ± 2.85 nm and 5.4 ± 0.75 nm, respectively. The penetration rate for cells treated with targeted nanoparticles was shown to be more than that of non-targeted nanoparticles. Moreover, it was revealed that the treatment of cells with ≥ 50 µg/ml FA-PG-SPIONs in combination with hyperthermia induced cytotoxicity in comparison to control cells. The results also showed that increasing the concentrations of targeted nanoparticles (FA-PG-SPIONs) and heating time would increase the value of thermal enhancement factor (TEF). In contrast, TEF values were not increased with increasing heating time and concentrations of non-targeted nanoparticles (PG-SPIONs). On the other hand, TEF values were increased with increasing concentrations and heating time so that the maximum TEF value was obtained at the highest concentration (FA-PG-SPION, 200 µg/ml) as well as the longest heating duration (60 min). Thus, it is concluded that FA-PG-SPIONs with concentrations ≥ 100 µg/ml could be introduced and used as hyperthermia sensitizing agents leading to enhanced cancer therapy efficiency.

N-(2'-Hydroxyphenyl)-2-propylpentanamide (OH-VPA), a histone deacetylase inhibitor, induces the release of nuclear HMGB1 and modifies ROS levels in HeLa cells.

N-(2′-Hydroxyphenyl)-2-propylpentanamide (OH-VPA) is a valproic acid (VPA) derivative with improved antiproliferative activity toward breast cancer (MCF-7, MDA-MB-231, and SKBr3) and human cervical cancer cell lines (HeLa) compared to that of VPA. However, the pharmacological mechanism of OH-VPA activity remains unknown. High-mobility group box 1 (HMGB1) is an important enzyme that is highly expressed in tumor cells and has a subcellular localization that is dependent on its acetylation or oxidative state. Therefore, in this study, we analyzed changes in HMGB1 sub-cellular localization and reactive oxygen species (ROS) as well as changes in HeLa cell viability in response to treatment with various concentrations of OH-VPA. This compound is formed by the covalent bond coupling VPA to a phenol group, which is capable of acting as a free radical scavenger due to its chemical similarities to quercetin. Our results show that OH-VPA induces nuclear to cytoplasmic translocation of HMGB1, as demonstrated by confocal microscopy observations and infrared spectra that revealed high quantities of acetylated HMGB1 in HeLa cells. Cells treated with 0.8 mM OH-VA exhibited decreased viability and increased ROS levels compared with the lower OH-VPA concentrations tested. Therefore, the antiproliferative mechanism of OH-VPA may be related to histone deacetylase (HDAC) inhibition, as is the case for VPA, which promotes high HMBG1 acetylation, which alters its subcellular localization. In addition, OH-VPA generates an imbalance in cellular ROS levels due to its biochemical activity.

Metamizole (dipyrone) – cytotoxic and antiproliferative effects on HeLa, HT-29 and MCF-7 cancer cell lines

Cancer pain treatment is a big challenge for healthcare providers and patients as well. The wide range of non-steroidal anti-inflammatory drugs (NSAIDs) used as painkillers in cancer patients, requires in-depth characterization of their effect on the disease process. The effects of NSAIDs have been widely studied over the last decades as preventive drugs in some oncological diseases. Metamizole is an NSAID belonging to the non-narcotic analgesics group and is highly recommended in oncology either alone or in combinations with opioid analgesics. There is a dearth of information regarding the cytotoxicity profile of metamizole and hence the present study evaluated the potential anticancer activity of metamizole in some permanent human tumour cell lines: HeLa, human cervical cancer cells; HT-29, a human colorectal adenocarcinoma cell line; and МСF-7, human breast adenocarcinoma cells. The studied tumour cells were sensitive to metamizole at doses higher than 25 μg/mL. Metamizole induced a statistically significant decrease in the viability of HeLa, HT-29 and MCF-7 cells in in vitro tests as measured by the MTT assay; the highest effect was observed at the 48th hour of the treatment. Metamizole could induce cell death by apoptosis. Metamizole also suppressed the migration of the three tumour cell lines. This was most clearly pronounced in HeLa cells. The results obtained indicate that metamizole is a suitable choice for the treatment of cancer pain and has prospects for further in-depth studies.

Synthesis and Properties of a New Thermo-sensitive Random Polyurethane with Tunable LCSTs

In this paper, a new class of thermo-sensitive random polyurethanes (PGLDs) have been prepared by polymerization of L-lysineethyl ester diisocyanate with five homologous derivatives of ethylene glycols. The structure and properties of polyurethanes were characterized by various techniques. The molecular weight Mn and structure of polyurethanes are found to be particularly significant factors to their thermo-sensitivity. Four polyurethanes except poly(L-lysine ethyl ester diisocyanate-co-ethylene glycol) show a reversible phase transition at the lower critical solution temperature, which can be easily tuned in the range of 4–35°C by combination of the monomers. Unexpectedly, the increasing number of –CH2CH2O–units in the main-chain of polyurethanes apparently acts as the hydrophobicity in the solution. The catalyst-free-synthesized polyurethanes with Mn of about (1–2) × 104 g/mol responses to the external temperature. Stannous octoate-catalysted polyurethanes with Mn of (3–6) × 104 g/mol exhibit no thermo-sensitivity because of their strong intra/intermolecular interactions. In addition, the viability of HeLa cells in 0.01–100 μg/mL solution reached to 80% after 24 and 48 h of incubation, indicating no cytotoxicity for polyurethanes.

Biological Activity of Flavonoids and Rare Sesquiterpene Lactones Isolated From Centaurea ragusina L.

The endemic Croatian species Centaurea ragusina L., like other species from the genus Centaurea, has been traditionally used in Croatia as an antibacterial agent and for the treatment of gastrointestinal and urogenital disorders. In several chromatographic steps, three flavonoids and three sesquiterpene lactones (STLs) were isolated and identified from the most active fractions of the ethanol extract. Two STLs, one for which we created the trivial name ragusinin, and hemistepsin A are here reported for the first time as constituents of the genus Centaurea. All six compounds were screened for their effect on several tumor and one normal cell lines. Among them, ragusinin showed the best bioactivity and high specificity to affect tumor murine SCCVII, human HeLa and Caco-2 cell lines, but not the viability of normal V79 fibroblasts. Due to these characteristics the action of ragusinin was investigated in more detail. Since DNA is the primary target for many drugs with antibacterial and anticancer activity, we studied its interaction with ragusinin. Rather moderate binding affinity to DNA excluded it as the primary target of ragusinin. Due to the possibility of STL interaction with glutathione (GSH), the ubiquitous peptide that traps reactive compounds and other xenobiotics to prevent damage to vital proteins and nucleic acids, its role in deactivation of ragusinin was evaluated. Addition of the GSH precursor N-acetyl-cysteine potentiated the viability of HeLa cells, while the addition of GSH inhibitor L-buthionine sulfoximine decreased it. Moreover, pre-treatment of HeLa cells with the inhibitor of glutathione-S-transferase decreased their viability indicating the detoxifying role of GSH in ragusinin treated cells. Cell death, derived by an accumulation of cells in a G2 phase of the cell cylce, was shown to be independent of poly (ADP-ribose) polymerase and caspase-3 cleavage pointing toward an alternative cell death pathway.

2-(2-nitrobenzylidene) indolin-3-one compound inhibits transmembrane prostate androgen-induced protein (TMEPAI) expression and cancer cell proliferation.

The transmembrane prostate androgen-induced protein (TMEPAI) is aberrantly expressed in many cancer and plays a crucial role in tumourigenesis, which makes it a potential cancer therapeutic target for drug discovery. Here, we employed a firefly luciferase reporter driven by the TMEPAI gene promoter to screen for compound capable of inhibiting the expression of TMEPAI, and the effects of TMEPAI inhibitor on cancer cell proliferation were evaluated using the colony formation assay, cell cycle analysis, Ki-67 immunofluorescence assay and EdU incorporation assay. 2-(2-nitrobenzylidene) indolin-3-one (JHY-A007-50) was identified and shown to effectively inhibit the TMEPAI promoter activity. Further studies revealed that JHY-A007-50 specifically inhibited the expression of TMEPAI at both the mRNA and protein levels. Moreover, we found that JHY-A007-50 could inhibit cell proliferation and induce cell cycle arrest at the G1 phase. Our results showed that overexpression of TMEPAI decreased the inhibitory effects of JHY-A007-50 on cancer cell proliferation, and JHY-A007-50 did not affect the cell viability of HeLa cells knocked down of TMEPAI. Taken together, these results suggest that compound JHY-A007-50 mediates the downregulation of TMEPAI expression and inhibits cell proliferation in cancer cells.

Angelica gigas Nakai Has Synergetic Effects on Doxorubicin-Induced Apoptosis.

Natural products are valuable sources for drug discovery because they have a wide variety of useful chemical components and biological properties. A quick reevaluation of the potential therapeutic properties of established natural products was made possible by the recent development of the methodology and improvement in the accuracy of an automated high-throughput screening system. In this study, we screened natural product libraries to detect compounds with anticancer effects using HeLa cells. Of the 420 plant extracts screened, the extract of Angelica gigas Nakai (AGN) was the most effective in reducing cell viability of HeLa cells. Markers of apoptosis, such as exposure of phosphatidylserine and cleavage of caspase-7 and PARP, were increased by treatment with the AGN extract. Treatment of the AGN extract increased expression of PKR as well as ATF4 and CHOP, the unfolded protein response genes. In addition, cotreatment of doxorubicin and the AGN extract significantly increased doxorubicin-induced apoptosis in HeLa cells. Decursin and decursinol angelate, which were known to have anticancer effects, were the main components of the AGN extract. These results suggest that the extract of AGN containing, decursin and decursinol angelate, increases doxorubicin susceptibility.

Extract of Penicillium sclerotiorum an endophytic fungus isolated from Cassia fistula L. induces cell cycle arrest leading to apoptosis through mitochondrial membrane depolarization in human cervical cancer cells.

Seventeen endophytic fungi were isolated from various tissues of Cassia fistula and the ethyl acetate extracts obtained from 21-day cultures of all the endophytic fungal isolates were initially screened for their cytotoxicity against HeLa (human cervical carcinoma) cells using MTT assay. Of these, Penicillium sclerotiorum extract (PSE), significantly affected the viability of HeLa cells in a dose-dependent manner. The extract of P. Sclerotiorum was further analyzed by GC-MS, which showed three compounds, hexadecanoic acid, oleic acid and benzoic acid to be the major active principles in the extracts.The extract was further tested for invitro cytotoxicity against five cancer cell lines. Of the cell lines tested, HeLa cells showed maximum sensitivity followed by A549, while A431 and U251 were moderately sensitive and MCF-7 was insensitive to the treatment. In addition, normal human embryonic kidney cells, HEK293 remained insensitive to the treatment. Furthermore, the mechanism of cytotoxic activity exhibited by PSE was investigated by evaluating cell cycle progression and apoptotic induction in HeLa cells. Cell cycle analysis revealed that the PSE arrested cells at S and G2/M phase of the cell cycle in a dose-dependent manner. Annexin V- Propidium iodide double staining showed that, the extract potentiates apoptosis rather than necrosis in cells. This was supported by the down regulation in the proapoptotic protein BCL2 and up regulation of BAX (BCL2 Associated X), tumor suppressor protein, p53 and Apaf-1 [Apoptotic Peptidase Activating Factor 1]. Loss of mitochondrial membrane potential and a distinct DNA fragmentation pattern observed following the treatment, suggest that the PSE treatment leads to activation of mitochondrial pathway of apoptosis. Further, the extract also exhibited both antioxidant and anti-angiogenic properties. These results indicate that endophytic fungi isolated from medicinal plants may serve as potential sources of the anti-cancerous compounds.

Antineoplastic activity of mitomycin C formulated in nanoemulsions-based essential oils on HeLa cervical cancer cells

Combining the essential oils (ESSOs) with the chemotherapeutic agent, mitomycin C (MMC), in nanoparticle can be beneficial in cancer therapy. The aim of the current study was to in vitro evaluate the antineoplastic effect of MMC, formulated in two different nanoemulsions (NE) based on two ESSOs, chamomile (Ch) and garlic (Gar), on HeLa cervical cancer cells. The z-average diameter of Ch-NE has slightly increased from 83.39 ± 12.85 nm to 91.18 ± 5.79 nm when mixed with MMC (Ch-MMC) whereas the z-average diameter of Gar-NE has markedly increased from 50.6 ± 1.96 nm to 75.64 ± 7.13 nm when loaded with MMC (Gar-MMC). The zeta potentials of both of Ch-NE and Ch-MMC, which were −1.91 ± 4.38 mV and −5.44 ± 5.26 mV, respectively, have differed from Gar-NE and Gar-MMC, which were 11.4 ± 2.29 mV and 11.5 ± 2.28 mV, respectively. Compared to MMC solution, the cell viabilities of HeLa cells, measured by the MTT assay, were reduced 42 and 20 times when subjected into Ch-MMC and Gar-MMC, respectively. The light microscopy images revealed that the cell membrane of the HeLa cells treated with Gar-NE or Gar-MMC were more altered relative to the cells treated with Ch-NE or Ch-MMC. In contrast, the nuclei of the HeLa cells, stained with DAPI and treated with Ch-NE or Ch-MMC, were more fragmented than the cells treated with Gar-NE or Gar-MMC, indicating that both of Ch-NE and Ch-MMC have passed the cell membrane and affected the nucleus directly whereas Gar-NE and Gar-MMC have got attached to the cell membrane causing damage to the cell. In conclusion, combining MMC with NE-based ESSOs has increased the cytotoxic effect of the MMC on the HeLa cells with different mechanism of actions.

Correlation between Electrical Field Strength and Pulse Width Analysis on Cell Viability

Electric field pulses can make living cell membrane electroporated either reversibly or irreversibly. This study determined the effects of electric field pulses on HeLa cancer cell viability. First set of experiment-Pulses electric fields (control, 200V/cm, 400V/cm, 600V/cm, 800V/cm, and 1000V/cm) with fixed pulse width (200µs) were performed on HeLa cells. Second set of experiment - Pulses width of (control,100µs, 500µs,1ms,5ms,10ms) with constant electric field strength of (600V/cm) were performed on HeLa cells. Number of pulses fixed to 1 for both sets of experiments. The quantitative determinations of cell viability measurement were performed by the aid of haemocytometer and trypan blue staining. The data shows that HeLa cell viability decreases as the electric field strength and pulse width. This is because electric pulses alter the cell’s transmembrane potential, causing disruption of the lipid bilayer, and increase the cell membrane permeability. Efficient electroporation outcome only will be gained by fine tuning the appropriate electric field parameter according to the application needed.

Amiodarone's major metabolite, desethylamiodarone, induces apoptosis in human cervical cancer cells.

Previously, we found that desethylamiodarone (DEA) may have therapeutic potentiality in bladder cancer. In this study, we determined its effects on human cervical cancer cells (HeLa). Cell viability was evaluated by Muse Cell Count & Viability Assay; cell apoptosis was detected by Muse Annexin V & Dead Cell Assay. Cell cycle was flow cytometrically determined by Muse Cell Cycle Kit and the morphological changes of the cells were observed under a fluorescence microscope after Hoechst 33342 staining. The changes in the expression levels of apoptosis-related proteins in the HeLa cells were assessed by immunoblot. Our results showed that DEA significantly inhibited the proliferation and viability of HeLa cells and induced apoptosis in vitro in dose-dependent and also in cell cycle-dependent manner because DEA induced G0/G1 phase arrest in the HeLa cell line. We found that DEA treatment downregulated the expression of phospho-Akt and phospho-Bad. In addition, DEA could downregulate expression of Bcl-2, upregulate Bax, and induce cytochrome c release. Our results indicate that DEA might have significance as an anti-tumor agent against human cervical cancer.

Anticancer effects of a novel chroman analog in HeLa cells are associated with G2‑phase arrest and mitochondrial‑mediated apoptosis.

In the present study, the anticancer activity of 1‑[(3S,4R)‑2,2‑dimethyl‑3‑oxo‑4‑(2‑piperidonyl)chroman‑6‑yl]‑3‑phenylurea (S32) was investigated by testing its effect in vitro on the growth of HeLa cells. First, we showed that the IC50 value of S32 was ~70 µM by using WST‑8 assay, and that it significantly inhibited the proliferation and viability of HeLa cells in a dose‑dependent manner after 48h. Morphological changes in apoptotic cells included cellular shrinkage and nuclear condensation. The results of [3H]‑thymidine incorporation and flow cytometric analysis indicated that S32 induced inhibition of DNA replication and G2‑phase cell cycle arrest. Moreover, S32 induced the levels of reactive oxygen species (ROS) and decreased the mitochondrial membrane potential (MMP) in a time‑dependent manner. Using Annexin V‑FITC/propidium iodide (PI) dual staining assay, we found that S32 noticeably increased early apoptosis in HeLa cells in a time‑dependent manner. The result of western blot analysis showed that the apoptotic induction was associated with an increase in Bax levels and a decrease in Bcl‑2 levels, which led to activation of caspase‑8, ‑9and‑3. Taken together, our findings demonstrated that S32 induces mitochondrial‑mediated apoptosis in HeLa cells and suggest that S32 has potential as an anticancer drug.

Amidated Scolopin‐2 inhibits proliferation and induces apoptosis of Hela cells in vitro and in vivo

This study aimed to investigate the effect of Scolopin-2, a cationic antimicrobial peptide from centipede venoms, and amidated Scolopin-2 on Hela cell viability in vitro and in vivo. The cellular proliferation was investigated with the MTT assay. Confocal laser scanning, flow cytometry, and Western blot analysis were employed to localize Scolopin-2-NH2 in Hela cells and to study the caused cells apoptosis. We subcutaneously injected Hela cells into BALB/c nude mice and studied if Scolopin-2-NH2 suppressed tumor growth in the mice. Scolopin-2-NH2 inhibited Hela proliferation in vitro in a dose-dependent manner with an IC50 of 35 μM. In addition, Scolopin-2-NH2 combined with mitochondria and regulated caspase-related apoptosis pathways in Hela cells. Scolopin-2-NH2 significantly suppressed tumor growth in the tumor-bearing mice without side effects, such as weight loss or abnormal changes in tissues, including liver, spleen, kidney, and lung. These results indicate Scolopin-2-NH2 may be a good therapeutic candidate for the treatment of Hela cervical cancer.

Effect of allyl isothiocyanate on the viability and apoptosis of the human cervical cancer HeLa cell line in vitro.

The present study aimed to investigate the effect of allyl isothiocyanate (AITC) on the viability and apoptosis of the human cervical cancer HeLa cell line in vitro, and to explore the potential underlying mechanisms of this. HeLa cells were treated with varying concentrations of AITC for different durations. The cell viability was then measured using a Cell Counting kit-8 assay and the apoptosis rate of the cells was detected using flow cytometry. Additionally, the B cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) mRNA expression levels were determined by reverse transcription-quantitative polymerase chain reaction, while the Bax and Bcl-2 protein expression levels in cells were detected by western blot analysis. AITC was revealed to inhibit the viability of HeLa cells. AITC was revealed to induce the apoptosis of HeLa cells, as the apoptosis rate increased gradually with an increase in the dose. As the concentration of AITC increased, the Bax mRNA expression level increased, whilst the Bcl-2 mRNA expression level decreased. Furthermore, the Bax protein expression intensity increased whilst Bcl-2 protein expression intensity decreased, thereby resulting in a decrease in the ratio of Bcl-2/Bax proteins. AITC may inhibit cell viability by inducing the apoptosis of HeLa cells and this may be accounted for by the imbalance in the Bcl-2/Bax expression ratio.

The Role of Fruits on the Cancer Incidence in Cusco Peru

Breast cancer remains as one of the most prevalent types of cancer in the world. Although Western industrialized nations rely on a contemporary approach to modern medicine, many developing countries, such as Peru, rely on traditional medicine and alternative natural remedies. Therefore, the focus of this study was to determine the extent to which natural remedies and nutrition, contribute to the prevention and incidence of cancer. Here, we focus on the use of certain fruits, like papaya and the noni fruit, as natural remedies for illness. First, survey data was collected in Cusco, Peru regarding which fruits are consumed as natural remedies for illness. Then, molecular techniques such as MTT assay and LDH Cytotoxicity assay were used to test the effect these fruits have on HeLa cells. The survey results showed, papaya and noni as the most commonly used fruits for illness. Noni powder and papaya fruit were extracted with ethyl acetate and ethanol and used in MTT and cytotoxicity assays. Preliminary results in mammalian cells showed no effect of the ethyl acetate extracts on the viability of HeLa cells. However, MTT assay results for the ethanol extraction of papaya skin showed a decrease in mammalian cell viability at high concentrations. Further experiments will be conducted to test fresh noni and papaya as well as other commonly eaten fruits in Peru for their anticancer properties.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The reducibility of heLa cell viability by Sargassum polycystum extracts

Cervical cancer is the second largest cause of death-related cancer in women. The efficacy of cancer drugs is still low. Bioactive of brown seaweed has been studied by in vitro and in vivo as anticancer. The aim of this study was to evaluate the cytotoxicity of Sargassum polycystum extracts on HeLa cell, to recognize bioactive on extract and estimate the interaction between the bioactive and target protein. S. polycystum was found from Talango Island waters and HeLa cell was obtained from Indonesian Science Institute. Sample was extracted by ethanol, ethyl acetate and hexane, concentrated and finally, extracts were assayed on HeLa cell. The viability of this cell was quantified on ELISA-Reader. The bioactive compounds of the extract were elucidated by GC-MS. The interaction between bioactive and target protein was evaluated by using in silico method. The result showed that the lowest viability of HeLa cell on n-hexane extracts treatment. The n-hexane extract of this seaweed contained benzenepropanoic acid. This compound reduced HeLa cell viability by reducing of thrombin concentration. In conclusion, the benzene propanoic acid of S. polycystum was the cytotoxic agent and it is potential agent for anti-cervical cancer.

Lupeol induces S-phase arrest and mitochondria-mediated apoptosis in cervical cancer cells

Cervical cancer is fourth most common fatal cancer in women worldwide. Lupeol is a dietary triterpenoid and has shown its anticancer efficacy against various cancer types with selectivity in targeting cancer cells. In the present study, anticancer efficacy and mechanism of action of a phytochemical, lupeol, in human cervical carcinoma (HeLa) cells has been examined. The anticancer efficacy of lupeol was assessed by trypan blue cell counting, annexin Vassay, cell cycle analysis, expression of apoptotic proteins by RT-PCR and Western blotting and assessment of mitochondrial ROS generation by mitosox and mitotracker assays. Our results demonstrated that lupeol decreased cell proliferation and viability of HeLa cells significantly (p less than 0.001). Lupeol induced S-phase cell cycle arrest and also decreased the expression of S-phase Cyclins and CDKs and increased the expression of cyclin-dependent kinase inhibitors, p21 at transcriptional and translational level. Further, lupeol induced apoptosis and increased the expression of apoptosis markers such as cleaved PARP and Bax:Bcl-2 ratio. Furthermore, mitosox and mitotracker dye incubation followed by FACS analysis showed an increase in mitochondrial superoxide generation and reduction in healthy mitochondrial mass. These results suggest that lupeol could be an effective chemotherapeutic agent against cervical carcinoma due to its growth inhibitory activity through induction of S-phase cell cycle arrest and apoptosis.

ε-Poly-l-Lysine/plasmid DNA nanoplexes for efficient gene delivery in vivo

The present work addresses the development and characterization of ε-Poly-l-Lysine/pDNA polyplexes and evaluation for their improved transfection efficacy and safety as compared to polyplexes prepared using Poly-l-Lysine and SuperFect®. Self-assembling polyplexes were prepared by varying the N/P ratio to obtain the optimum size, a net positive zeta potential and gel retardation. The stability in presence of DNase I and serum was assured using gel retardation assay. Their appreciable uptake in MCF-7 and 3.5, 3.79 and 4.79-fold higher transfection compared to PLL/pDNA polyplexes and 1.60, 1.53 and 1.79-fold higher transfection compared to SuperFect®/pDNA polyplexes in MCF-7, HeLa and HEK-293 cell lines respectively, affirmed the enhanced transfection of ε-PLL/pDNA polyplexes which was well supported with in vivo transfection and gene expression studies. The <8% in vitro hemolysis and >98% viability of MCF-7, HeLa and HEK-293 cells in presence of ε-PLL/pDNA polyplexes addressed their safety, which was also ensured using in vivo toxicity studies, where hemocompatibility, unaltered levels of biochemical markers and histology of vital organs confirmed ε-PLL to be an effective and safer alternative for non-viral genetic vectors.

Sesamin induces ER stress-mediated apoptosis and activates autophagy in cervical cancer cells

AimsSesamin, a major lignan of sesame oil, has demonstrated anticancer properties. However, its anticancer effects on cervical cancer have not been studied. Here, we investigated the effects of sesamin on cervical cancer (HeLa) cell line and explored the underlying mechanisms. Main methodsHeLa cells were cultured with sesamin. CCK-8 and scratch wound test were applied to detect the proliferation and migration ability, while flow cytometry and TUNEL staining were applied to detect apoptosis. The expression of Bax and Bcl-2 was assessed by Western blotting. Further observe the ultrastructure using transmission electron microscopy (TEM) and detect the expression of caspase-12, GRP78, GADD153, IRE1α, p-IRE1α, JNK, p-JNK, LC3I/II and beclin-1. In addition, HeLa cells were treated with 3-MA (an autophagy inhibitor) and/or sesamin. Then detect the expression of LC3I/II and cell viability. Key findingsCCK-8 and scratch wound test revealed that sesamin inhibits HeLa cells proliferation and migration, while flow cytometry and TUNEL staining indicated that sesamin induces apoptosis in these cells. In sesamin group, the expression of Bax, caspase-12, GRP78, GADD153, p-IRE1α, p-JNK, LC3I/II and beclin-1 was up-regulated while Bcl-2 was down-regulated compared to control group. Further research revealed that sesamin also induces Hela cells autophagy and inhibition of autophagy increases cell viability of sesamin-treated HeLa cells. SignificanceSesamin inhibits proliferation/migration of HeLa cells and induces ER stress-mediated apoptosis through IRE1α/JNK pathway, and that it activates autophagy and autophagic death in these cells, further validate the anticancer effect of sesamin.