MATERIAL AND METHOD

To study and estimate the most prevailing form of cancer in all over the world,

The data is collected from institute and websites;

.National Institute center.

.American society for cancer.

.World health organization.

Since1983, instance cases of cancer in all over the world have been reported by hospital
departments, pathology laboratories and practising physicians, mainly dermatologists.

Only one skin cancer of each morphologic type is recorded most prevailing form of cancer in
New Zealand , with an indication in the register of persons with multiple skin cancers of a given
type located to more than one site of the body. Incidence rates are therefore based on the number
of persons with tumors rather than the number of tumors. All cases of malignant melanoma,
basal cell carcinoma and squamous cell carcinoma of the skin identify in the period January 1989
to march 2011. Rates were calculated as average annual incidence rates per 100,000 persons with
the Worldwide population on 1 march 2018 as the denominator. The incidence rates have been
age-standardized to the World Population by the direct method.

RNA isolation from plasma and qRT-PCR

To detect the gastric cancer, blood samples, which were stored in the separate vacuum cubes,
were centrifuged at 3,000 rpm for 10 min at 4°C. The supernatant was collected and stored at -
80°C for qRT-PCR. According to QIAGEN miRNeasy Mini Kit (Qiagen, Valencia, CA, USA),
the RNA was extracted from 100 μL of plasma. CDNA of miR-217 was conducted through the
instructions of the kit.

Biomarkers for early detection:

These biomarkers are based mainly on inherited or somatically acquired susceptibilities, in the
form of altered genes such as MSH2 and MLH in hereditary non-polyposis colorectal cancer,
PRB in hereditary retinoblastoma, and BRCA1 and BRCA2 mutated genes that predispose to
breast cancer. Theoretically, they could provide the opportunity to intervene during the natural
progression of the cancer, to cause inhibition, regression, or even elimination of the disease.

Proteomics

Proteomic methods detect the functioning units of expressed genes,34 through biochemical
analysis of cellular proteins, to provide a protein fingerprint.35 The proteome reflects both the
intrinsic genetic programme of the cell and the impact of its immediate environment and is
therefore valuable in biomarker discovery. Distinct changes that occur at the protein level during
the transformation of a normal cell into a neoplastic cell include altered expression, differential
protein modification, changes in specific activity, and inappropriate localisation, all of which
may affect cellular function.

Skin cancer can be detected by using following techniques. Human endothelial cells (strain
H3605) were from J. Wessendorf and T. Maciag (American Red Cross, Rockville, MD) (20),
neonatal melanocytes were from Z. Abdel-Malek (University of Cincinnati), adult melanocytes
were from shave biopsies , mammary cells were from V. Band (New England Medical Center,
Boston) and ovarian cells were from N. Auersperg (23). SV40-WI38, C33a, U20S, SAOS and
HTB9 were from the American Type Culture Collection.

[3H]Thymidine Labeling

Sparse cells (1-5 x 103 per cm2) were given 10 ,uCi of [3H]thymidine (60-80 Ci/mmol; 1 Ci =
37 GBq) per ml for 48-72 hr, stained where indicated, washed in phosphate-buffered saline
(PBS), rinsed twice in methanol and processed for autoradiography.

Beta-Galactosidase Staining

Cells were washed in PBS, fixed for 3-5 min (room temperature) in 2% formaldehyde/ 0.2%
glutaraldehyde (or 3% formaldehyde), washed and incubated at 37°C (no C02) with fresh
senescence associated (3-Gal (SA-,3-Gal) stain solution: 1 mg of 5-bromo- 4-chloro-3-indolyl
P3-D-galactoside (X-Gal) per ml (stock = 20 mg of dimethyl formamide per ml)/40 mM citric
acid/sodium phosphate, pH 6.0/5 mM potassium ferrocyanide/5 mM potassium ferricyanide/150
mM NaCl/2 mM MgCl2. Staining was evident in 2-4 hr and maximal in 12-16 hr. To detect
lysosomal ,B-Gal, the citric acid/sodium phosphate was pH 4.0.

Skin Samples

Human skin from individuals undergoing Moh's micrographic surgery for skin cancer was
rapidly frozen in liquid nitrogen, and mounted in OCT. Thin sections (4 ,um) were cut, mounted
onto glass slides, fixed in 1% formalin in PBS for 1 min at room temperature, washed in PBS,
immersed overnight in SA-3-Gal staining solution, counterstained with eosin, and viewed under
bright field at 100-200X magnification.