Expression of CCND1, P16 and CDK6 in Human Basal Cell Carcinoma

Basal cell carcinoma (BCC) is the most common cancer among skin cancers. The incidence of cutaneous malignant melanoma (CMM) and non-melanoma skin cancer (NMSC) has increased more than 600% worldwide since the 1940s. Carcinogenesis is a multi-step process involving multiple genetic alterations. The connection between cell cycle proliferation and cancer resulting in deregulated cellular proliferation leads to cancer. Cancer has been associated with disturbances in cell cycle regulation. Recent studies have shown that p16, CDK6 and CCND1 mRNA genes and protein expression are involved in the tumorgenesis of skin cancer. These genes play a role in cell cycle proliferation. In this study, we assessed the expression of a cyclin, a cyclin dependent kinase, and a cyclin kinase inhibitor in skin BCC tissue. Reverse Transcription in situ polymerase chain reaction (RT in situ PCR) and immunohistochemistry (IHC) were used to detect the expression of p16, CDK6 and CCND1 mRNA genes through them of protein expression in human skin BCC tissue The results show that p16, CDK6 and CCND1 mRNA genes and protein are expressed in both normal and human skin BCC tissues. CCND1, CDK6 and p16 mRNA can be found to be expressed mostly in cytoplasm. The mRNA expression in BCC is higher than normal skin tissue. Protein expression of CCND1 and p16 in different BCC tissue are greater than normal skin tissue. p16 mRNA and protein expression is stronger than other genes. RT in situ PCR and IHC analysis data showed significant expression of CCND1, p16 mRNA and protein in BCC compared to normal skin tissue (p<0.05%). Investigation on Iranian samples showed the protein expression of CDK6 is not significant (p>0.05%) but the expression of mRNA for CDK6 gene is significant (p<0.05%). The findings of IHC study on tissue microarray (TMA) demonstrated significant protein expression of p16 and CCND1 genes (p<0.05%) which support findings on Iranian samples. Taken together, these data provide evidence that cell cycle deregulation in G1-phase is a critical event during the course of carcinogenesis of BCC. In conclusion, this study showed that p16, CDK6 and CCND1 are involved in the process of tumorgenesis in human BCC. p16, CDK6 and CCND1 mRNA genes are expressed to induce cell cycle proliferation and also the protein expression of these genes can influence proliferation of the cell cycle. RT in situ PCR study on ten Iranian samples illustrated significant expression of p16 (p=0.026), CDK6 (p=0.015) and CCND1 (p=0.021) mRNA genes (p<0.05%). There is a direct correlation between p16 and CCND1 and also between p16 and CDK6. There is no correlation between CCND1 and CDK6. IHC analysis on the Iranian samples demonstrated significant protein expression of p16 (p=0.019) and CCND1 (p=0.021) (p<0.05%) but CDK6 protein expression is not significant (p=0.082). Direct correlation between p16 and CCND1 was obtained. TMA samples were used for the IHC study only for p16 (p=0.008) and CCND1 (p=0.024) due to insufficient tissue to perform complete study with IHC. Even CDK6 could not be done because of insufficient samples then the result of TMA samples using IHC supports the findings on the Iranian samples about protein expression of CCND1 and p16 genes. RT in situ PCR is a sensitive method to study specific mRNA genes. However there are problems in getting good results as well as their interpretation. IHC on the other hand shows more reliable results. These methods may be used in the clinical setting and as it can be used to predict tumor behavior including cellular proliferation which can affect the mode of therapy

Delivery of recombinant plasmid containing human insulin gene-GLP1 promoter into L cells in the rats with a type-1 diabetes

The purpose of this study was to determine whether the treatment with recombinant plasmid consisted of human GLP1 promoter and insulin gene can treat diabetic rats. Rats were induced type-1 diabetes mellitus (T1DM) by a single dose of intraperitoneal injection of streptozotocin (STZ)at dose of 55mg/kg. The induction of diabetes was confirmed in rats by checking the blood glucose level for seven days. The recombinant plasmid, GLP1/Ins/pBud plasmid, was wrapped with chitosan and then transferred to diabetic rats by force feeding. The blood glucose level was checked from the tips of the tails by needle puncture using a glucometer and test strips. The blood levels of human and rat insulin were assessed by enzyme-linked immunosorbent assay (ELISA). The results showed no significant effects of orally treatment with recombinant plasmid DNA at both doses of 100 and 600 μg/mL of the human insulin level in diabetic rats (p>0.05). The human insulin level was significantly increased by orally treatment at dose of 300 μg/mL (p=0.04). The findings indicated that the intraperitoneal injection of 300 μg/mL of this nanoparticle complex prominently increased the human insulin level in diabetic rats in contrast to both doses of 100 and 600 μg/mL. Despite above results, both methods was not effective enough to decrease the blood glucose levels in diabetic rats. It was concluded that the treatment of diabetic rats with recombinant plasmid consisted of human GLP1 promoter and insulin gene was not effective to reduce the blood glucose levels in diabetic rats

Gene expression of CDK6 and CCND1 genes in basal cell carcinoma

Basal cell carcinoma (BCC) is the most common cancer among skin cancers. Cell cycle deregulation in G1-phase is a critical event during the course of carcinogenesis, which is probably much more important than other phases of cell cycle, during the course of skin carcinogenesis. CCND1 and CDK6 are important components of Retinoblastoma regulatory pathway in arrest and uncontrolled proliferation of cell cycle. To determine the expression pattern of CDK6, CCND1 in BCC, this study involved ten samples of paraffin embedded of BCC tissues. Two selected normal skin tissue were investigated using RT in situ PCR and Immunohistochemistry (IHC) techniques. Nuclear and cytoplasmic staining intensity of samples within tumor cells and normal tissue illustrated a different mRNA and protein expression. This study represents significant expression of CCND1 and CDK6 genes in BCC (alpha level is 0.05). CDK6 and CCND1 mRNA, and protein of these genes are expressed to induce the cell cycle proliferation and the influence proliferation of cell cycle and BCC

Correlation of cognitive impairment with constipation and renal failure

The improvement of health care support has greatly extended the average life expectancy over the last 50 years, which has increased the rate of cognitive decline consequently. The avoidance of risk factors such as toxins, stress and somatic diseases can be protective against the reduction of cognitive function in the elderly. This study aimed to determine the effects of socio-demographic factors, constipation and renal failure on cognitive status among 2322 samples who were the non-institutionalized Malaysian elderly. The multiple logistic regression analysis was applied to estimate the risk of such factors on cognitive decline in subjects. Approximately, 77.54% of samples experienced cognitive impairment. The results showed that advanced age (odds ratio (OR) = 1.03), Malay ethnic (OR = 2.15), constipation (OR = 3.31) and renal failure (OR= 4.42), significantly increased the risk of cognitive impairment in subjects (p<0.05). In addition, education (OR = 0.38) significantly reduced the risk. However, we concluded that age, Malay ethnic, constipation and renal failure increased the risk of cognitive impairment in subjects but education reduced the risk

Pridonosi li polimorfizam gena GSTP1 oštećenju genoma uzrokovanom starenjem i profesionalnom izloženosti?

The aim of our study was to see the effects of GSTP1 polymorphism on biomarkers of ageing, including micronuclei (MN), comet tail length, and relative telomere length in automobile repair workers, who are exposed to a broad spectrum of potential mutagens. The analysis was performed on buccal cells collected from occupationally exposed and non-exposed (control) subjects. Samples were analysed using cytogenetic and molecular methods, including restriction fragment length polymorphism (RFLP), MN test, comet assay, and real-time PCR. The results confi rmed the DNA damaging effects of substances used in the mechanical workshops, but did not confirm the infl uence of GSTP1 gene polymorphism on DNA damage. However, further studies on both occupationally exposed and control populations are needed to understand the relationship between GSTP1 polymorphism and genome damage.Na populaciji radnika zaposlenih u radionicama za popravak automobila koji su svakodnevno izloženi različitim vrstama potencijalnih mutagena istražili smo utjecaj polimorfizma gena GSTP1 na vrijednosti biomarkera starenja, ključujući pojavu mikronukleusa (MN), dužinu repa kometa te relativnu dužinu telomera. Analize su provedene na stanicama bukalne sluznice, skupljenim od izloženih ispitanika i odgovarajuće neizložene (kontrolne) populacije. Uzorci su analizirani primjenom citogenetičkih i molekularnobioloških metoda, uključujući polimorfi zam restrikcijskih fragmenata na osnovi njihove duljine (engl. restriction fragment length polymorphism, RFLP), MN-test, komet-test, i lančanu reakciju polimerazom u stvarnom vremenu (engl. real-time PCR). Dobiveni nalazi potvrđuju da izloženost radnika mutagenima oštećuje njihovu DNA, ali nisu pokazali da polimorfi zam gena GSTP1 značajno utječe na razinu oštećenja DNA. Zbog malog broja ispitanika uključenog u ovo istraživanje za bolje razumijevanje odnosa između polimorfi zma gena GSTP1 i oštećenja DNA potrebna su daljnja istraživanja, i na profesionalno izloženim ispitanicima i na ispitanicima kontrolne populacije

p16 gene expression in basal cell carcinoma.

Background: Basal cell carcinoma (BCC) develops predominantly in sun-exposed skin in fair-skinned individuals prone to sunburn. BCC typically occurs in adults. High exposure to ultraviolet (UV) radiation increases rate of developing BCC, a slowly growing tumor that occurs in hair-growing squamous epithelium and rarely metastasizes. In genetic studies, BCC patients have cell-cycle abnormalities of different parts of the signaling pathway. Retinoblastoma regulatory pathway is important in cell cycle arrest. In this pathway, p16INK4a, an inhibitor of Rb pathway, binds to CDK4 and CDK6 competitively with cyclin D1 to prevent phosphorylation of tumor suppressor pRB gene. Alteration of this pathway contributes to development of human cancers and also is effective in skin cancers. In this study, we analyzed mRNA expression using in situ RT-PCR and the role of immunohistochemical expression of p16INK4a in BCC. Methods: Expression of p16 in ten samples of Iranian paraffin-embedded skin BCC were studied using in situ RT-PCR and immunohistochemistry on p16INK4a gene. Results: Nuclear and cytoplasmic staining intensity of samples within tumor cells and normal skin tissue illustrates different mRNA and protein expression of p16 gene. mRNA of p16 gene and the expressed protein induce cell cycle proliferation and involve both tumor tissue as well as normal skin tissue. However, in this study it was found that there is significant protein and mRNA expression in BCC cells when compared to normal skin tissue (p <0.05). Conclusions: p16 gene is involved in the pathogenesis of human skin BCC in view of increased p16 mRNA and expressed protein within tumor cells

The effects of sleep quality, physical activity, and environmental quality on the risk of falls in dementia

OBJECTIVES: This study aimed to identify the effects of sleep quality, physical activity, environmental quality, age, ethnicity, sex differences, marital status, and educational level on the risk of falls in the elderly individuals with dementia. METHODOLOGY: Data were derived from a group of 1210 Malaysian elderly individuals who were noninstitutionalized and demented. The multiple logistic regression model was applied to estimate the risk of falls in respondents. RESULTS: Approximately the prevalence of falls was 17% among the individuals. The results of multiple logistic regression analysis revealed that age (odds ratio [OR] = 1.03), ethnicity (OR = 1.76), sleep quality (OR = 1.46), and environmental quality (OR = 0.62) significantly affected the risk of falls in individuals (P .05). CONCLUSION: It was found that age, ethnic non-Malay, and sleep disruption increased the risk of falls in respondents, but high environmental quality reduced the risk of falls

A research on functional status, environmental conditions and risk of falls in dementia

This study aimed to determine the effects of disability, physical activity, and functional status as well as environmental conditions on the risk of falls among the elderly with dementia after adjusting for sociodemographic factors. Data were derived from a group including 1210 Malaysian elderly who were demented and noninstitutionalized. The study was a national cross-sectional survey that was entitled “Determinants of Health Status among Older Malaysians.” Approximately 17% of subjects experienced falls. The results showed that ethnic non-Malay (OR = 1.73) and functional decline (OR = 1.67) significantly increased the risk of falls in samples (P 0.05). It was concluded that functional decline and ethnic non-Malay increased the risk of falls but the increased environmental quality reduced falls

Association between dentures and the rate of falls in dementia

Background: Poor oral health, chronic diseases, functional decline, and low cognitive ability can increase the risk of falls in the elderly. Objectives: The current study aimed to show the effects of oral health, diabetes mellitus (DM), hypertension (HT), heart disease, functional status, and sociodemographic factors on the risk of falls in elderly with dementia. Materials and methods: The sample comprised 1,210 Malaysian elderly who were demented and noninstitutionalized. This study was a national cross-sectional survey entitled “Determinants of Health Status among Older Malaysians”. The effects of age, ethnicity, sex differences, marital status, educational level, oral health, DM, HT, heart disease, and functional status on the risk of falls were evaluated. The multiple logistic regression model was used to estimate the effects of contributing variables on the risk of falls in samples. Results: The prevalence of falls was approximately 17% in subjects. It was found that age (odds ratio [OR] 1.02), non-Malay ethnicity (OR 1.66), heart disease (OR 1.92), and functional decline (OR 1.58) significantly increased the risk of falls in respondents (P<0.05). Furthermore, having teeth (OR 0.59) and dentures (OR 0.66) significantly decreased the rate of falls (P<0.05). Conclusion: It was concluded that age, non-Malay ethnicity, functional decline, heart disease, and oral health significantly affected falls in dementia