The antitumor activity of EGCG is attributed to its capacity to mediate signaling pathways and regulate cells’ undesired survival

and proliferation. The cell death by EGCG is initiated by the intrinsic pathways in various cancers (Table 1). EGCG suppresses
ERK1/2, NF-κB, and Akt-mediated signaling and activates p53 and PTEN/p21 for cell apoptosis along with the alteration of the
Bcl-2 protein ratio (Almatroodi et al., 2020). Induction of apoptosis takes place through some pathways which incorporate
intrinsic and extrinsic pathways, regulatory proteins, the strain on the endoplasmic reticulum through the activation of
caspase-mediated pathways, death receptors, downregulation of several anti-apoptotic proteins, upregulation of Bad, and Bax
pro-apoptotic proteins in human adrenal cancers cells (Wu et al., 2019) (Figure 2). The cell cycle arrest is triggered by EGCG at
G0/G1 phase and occurs via the regulation of several cyclins in pancreatic cancer cells. In cyclins, EGCG stops cyclin D1 and
turns on p21, which in addition consists of ERK, IKK, and PI3K signaling pathways in which colorectal cancer cells are being
inhibited from proliferating (Rady et al., 2018) (Figure 2). In cervical cancer, EGCG prevented the spread, invasion, and
migration of HeLa cells via down-and upregulation of MMP-9 and TIMP-1 genes, respectively (Sharma et al., 2012). EGCG
inhibited tumor growth by activating VEGF/VEGFR axis, interrupting the HIF-1a expression and other foremost growth factors,
inactivation of PAR2-AP, ERK1/2, and NF-κB pathways were blocked (Zhou et al., 2012). In esophageal tumor cells, EGCG
suppresses cellular viability through the reduction of p-ERK1/2, c-Jun, and COX-2, which in addition reasons the activation of
caspase-3 in conjunction with the suppression of the COX-2 expression (Ye et al., 2012). EGCG significantly suppresses the
glycolytic enzymes’ mRNA levels in breast cancer cells (Wei et al., 2018). In colorectal-cancer-cell, EGCG activates caspase-3 for
apoptosis, PARP, downregulation of STAT3 and phosphorylated STAT3 (p-STAT3), decreased Bcl-2 MCL-1, vimentin, along with
the increase in E-cadherin (Luo et al., 2021). EGCG set up its important function for inhibition of glioma and showed Mitogen-
activated protein kinase pathway (MAPK) involvement in apoptosis and proliferation (Figure 2). EGCG treatment with TRAIL
induces rapid apoptosis which could be a striking approach for treating several gliomas.

polyphenols display more capability than other compounds by blocking more than one target in pathways and mechanisms
associated with cancer progression. Various in-vitro studies suggest that the polyphenols modulate Nrf 2 and NF-kB activation
in cells and also influence the MAPK and PI3K function in the cells, thereby establishing a prominent role in cancer cell
progression. In connection, the possibility of combining conventional drugs with polyphenols (in individual or combination)
offers valuable advantages for developing more anti-cancer therapies with greater efficacy. Although, there is no doubt about
the role of oxidation in the occurrence of mutations in DNA and the increased risk of cancer, the isolated action of a particular
polyphenol or polyphenolic compounds, in general, is unlikely to have a distant effect on the gastrointestinal tract - for
example, in the mammary gland or lungs. While data on the beneficial effects of polyphenolic compounds as anticancer
agents, it has been observed that all these benefits depend not only on the food content of polyphenols, but especially on
their

Proteosome activity:

Proteasome activity in cancer : proteasome activity are of interest because its altered activity may
be associated with not only the pathobiology of cells but also therapeutic opportunities.

We have gone for A study , we also reported that a newly-developed proteasome inhibitor,
a disulfiram-copper complex, potently inhibits the proteasomal activity in cultured
breast cancer MDA-MB-231 and MCF7 cells before induction of apoptotic cell death. In
sharp contrast, the disulfiram-copper complex neither inhibited the proteasome nor
induced apoptosis in normal, immortalized MCF-7cells . Furthermore, MDA-MB-231
cells, containing copper concentrations similar to those found in patients, treated with
disulfiram, demonstrated proteasome-inhibitory and apoptosis inducing effects.
Likewise, in mice bearing MDA-MB-231 tumor xenografts, disulfiram significantly
inhibited the tumor growth associated with in vivo proteasome inhibition and apoptosis
induction. In this system, it appears that inhibition of the proteasomal activity can be
achieved by targeting tumor cellular copper with disulfiram to selectively induce
apoptosis in tumor cells.
Others have found similar, potent proteasomal inhibitory effects in cancer cells while
normal cells appear unaffected. Accumulation of NOXA (a pro-apoptotic member of the
Bcl-2 family of proteins) can negate the multiple Bcl-2 pro-survival family members and
facilitate mitochondrial cytochrome c, SMAC, and apoptosis-inducing factor (AIF) release
with subsequent DNA degradation and apoptosis. It has been shown that melanoma and
myeloma cells accumulate NOXA upon treatment with MG-132, lactacystin, and
bortezomib while NOXA induction was not triggered in untransformed melanocytes.
Furthermore, the proteasome inhibitor Z-Ile-Glu(OtBu)-Ala-Leucinal (PSI) has been
found to induce apoptosis in multiple human myeloid leukemia cell lines at low doses
(IC50 ranging from 5 to 25 nmol/l) indicated by nuclear DNA fragmentation, cleavage of
poly (ADP-ribose) polymerase (PARP) and of beta-catenin. Importantly, PSI was found
to be cytotoxic to leukemia, but not normal hematopoietic progenitor cells. These
studies indicate that the untransformed cell line is much less sensitive to proteasome
inhibitor-induced apoptosis.

These findings suggest that one possible explanation for the difference of tumor vs.
normal cells in response to proteasome inhibition is the basal proteasome activity level.
At least in the case of MCF 7 or MCF 10A or MDAMB 231, these tumor cells may require
higher proteasomal chymotrypsin-like activity for rapid proliferation compared to
normal cells.

Flurosence :

Here we have gone for Intracellular testing of MaBiDZ sensor. CLSM (confocal microscope) images of (A)
Twist-overexpressing MCF-7 cancer cells treated with MaBiDZ sensor with magnetic field applied and (C)
no magnetic field applied. Analogously treated cervical epithelial cells (expressing low levels of Twist) with
(B) MDAMB 231 magnetic field applied and (D) without magnetic field. Images were taken after 2.5 h of
incubation time. Nuclei are stained with Hoechst nuclear stain and visualized with 408 nm laser. Surfaces
are stained with anti-epithelial cell adhesion molecule (EpCAM) antibody and visualized with a 635 nm
laser. Fluorescence from the MaBiDZ probe is visualized with the 488 nm laser. Corresponding flow
cytometry data are shown as insets below each image. The gates on flow cytometry plots indicate percent
of EpCAM positive cells with low and high MaBiDZ fluorescence. The number of internalized particles was
estimated to be ca. 1 × 106 MaBiDZ per cell in the Scale bar is 20 μm.

To quantify the intracellular signaling of the MaBiDZ probe, we examined large population of cells
treated with probes using flow cytometry. This method eliminates variations that can be observed
using CLSM, which only permits the examination of a small fraction of cells. Flow cytometry
results  show that MCF-7 cells treated with MaBiDZ and a magnetic field (ON state) exhibited 4
times greater fluorescence than MaBiDZ-treated MCF-7 cells without a magnetic field (OFF state),
thus confirming the magnetic field-dependent switch-like effect of this system . When compared
to the control noncancerous HCX cells, MCF-7 cells exhibited a 20-fold fluorescence
enhancement. It is important to note that significant signaling was apparent after only 2.5 h, as
opposed to a previous technique that required an incubation of 12 h before a signal could be
detected. The fluorescence data from CLSM and flow cytometry measurements of whole cells
was validated using fluorescence data of cell lysates . This data was in good agreement with
measurements of Twist levels from whole cells.

Our next aim was to investigate the mechanisms that promote the observed signaling efficiency
and enhancement of MaBiDz within the cell. We hypothesized that the magnetic field plays a role
in enhancing cellular entry and intracellular transport kinetics, based on previous reports.  To
investigate this, we examined a small window of events upon cellular entry of MaBiDZ, both with
(ON) and without (OFF) a magnetic field. Previous studies show that nanoparticles enter cells by
endocytosis, and are subsequently either stored in endosomes or lysosomes, or undergo
endosomal escape. If these intracellular nanoparticles cannot escape from the endosome or
lysosome, they are not available for intracellular sensing. Therefore, we investigated the
distribution and colocalization of the oligo-modified MDAMB 231 Cells and MCF7 cells and
endosomes by CLSM at various time points. Results indicate that, at the peak of endosomal
internalization of MDAMB 231, the ON state demonstrated about 50% less colocalization of MaB
and endosomes compared to the OFF state. Though the mechanism is under investigation, the
data suggest that a magnetic field mitigates the bottleneck of endosomal sequestering, freeing
nanoparticles for sensing functions in the cytoplasm.

As a result we have designed a fluorescent hybridization MaBiDZ mRNA sensing system that can
be activated by a magnetic field at the desired time. MaBiDZ sensing technology produces low
background fluorescence that can be instantly activated by a magnetic field. We have already
demonstrated  the sensor which can be used for magnetic field-dependent mRNA sensing in living
cells. so we detection of specific mRNA in live cells within 2.5 h after applying a magnetic field,
which is a significant improvement in comparison with current techniques. We hope that the
MaBiDZ technology introduced here will add to the toolbox of techniques for RNA analysis in live
cells. The developed approach can find much broader applications than the presently
demonstrated cancer biomarker analysis example.