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ORIGINAL ARTICLE
» Similar in PUBMED Year : 2004 | Volume : 41 | Issue : 1 | Page : 25-31
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Patel PS Alterations in plasma lipid profile patterns in head and neck cancer and oral precancerous conditions
Shah MH
Jha FP Prabhudas S Patel, MH Shah, FP Jha, GN Raval, RM Rawal, MM Patel, JB Patel, DD Patel
Raval GN Biochemistry Research Division, Department of Cancer Biology; The Gujarat Cancer & Research Institute,
Rawal RM Asarwa, Ahmedabad - 380016, India
Patel MM
Patel JB
Patel DD Correspondence Address:
Prabhudas S Patel
» Search in Google Scholar Biochemistry Research Division, Department of Cancer Biology; The Gujarat Cancer & Research Institute,
for Asarwa, Ahmedabad - 380016
India
Patel PS
Shah MH PMID: 15105576
Jha FP
Raval GN
Rawal RM
Patel MM » Abstract
Patel JB
Patel DD BACKGROUND : The changes in lipid profile have long been associated with cancer because lipids play a key
role in maintenance of cell integrity. AIMS : The present study evaluated alterations in plasma lipid profile in
»Search keywords untreated head and neck cancer patients as well as patients with oral precancerous conditions (OPC) and its
association with habit of tobacco consumption. MATERIAL AND METHODS : This hospital-based case
Lipids control study included 184 head and neck cancer patients, 153 patients with OPC and 52 controls. Plasma lipids
Cholesterol including: (i) Total cholesterol, (ii) LDL cholesterol (LDLC), (iii) HDL cholesterol (HDLC) (iv) VLDL cholesterol
Triglycerides (VLDLC) and (v) triglycerides were analysed by spectrophotometric kits. STATISTICAL ANALYSIS USED :
Head and neck cancer Student's t-test was performed to compare mean values of the parameters. RESULTS : A significant decrease in
Oral precancerous plasma total cholesterol and HDLC was observed in cancer patients (P=0.008 and P=0.000 respectively) as well
conditions as in patients with OPC (P=0.014 and P=0.000, respectively) as compared to the controls. The plasma VLDL
and triglycerides levels were significantly lower in cancer patients as compared to the patients with OPC
» [PDF Not available] * (P=0.04) and controls (P=0.059). The tobacco habituates showed lower plasma lipid levels than the
» Citation Manager non-habituates. Our data strengthen the evidence of an inverse relationship between plasma lipid levels and head
» Access Statistics and neck malignancies as well as OPC. CONCLUSION :The lower levels of plasma cholesterol and other lipid
constituents in patients might be due to their increased utilization by neoplastic cells for new membrane
» Reader Comments
biogenesis. The findings strongly warrant an in-depth study of alterations in plasma lipid profile in head neck
» Email Alert * cancer patients.
» Add to My List *
* Registration required (free)

Keywords: Lipids, Cholesterol, Triglycerides, Head and neck cancer, Oral precancerous conditions
In this article
» Abstract
How to cite this article:
» Introduction
Patel PS, Shah MH, Jha FP, Raval GN, Rawal RM, Patel MM, Patel JB, Patel DD. Alterations in plasma lipid
» Material and Methods profile patterns in head and neck cancer and oral precancerous conditions. Indian J Cancer 2004;41:25-31
» Results
» Discussion
How to cite this URL:
» Acknowledgements Patel PS, Shah MH, Jha FP, Raval GN, Rawal RM, Patel MM, Patel JB, Patel DD. Alterations in plasma lipid

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Alterations in plasma lipid profile patterns in head and neck cancer and or... http://www.indianjcancer.com/article.asp?issn=0019-509X;year=2004;v...

» References profile patterns in head and neck cancer and oral precancerous conditions. Indian J Cancer [serial online] 2004
» Article Figures [cited 2010 Sep 6];41:25-31. Available from: http://www.indianjcancer.com/text.asp?2004/41/1/25/12341
» Article Tables

Article Access Statistics

» Introduction
Viewed 6879
Printed 50
Emailed 2 Lipids are major cell membrane components essential for various biological functions including cell growth and
PDF Downloaded 4 division of normal and malignant tissues. Usefulness of variations in tissue/blood cholesterol levels in diagnosis
Comments [Add] and treatment of various diseases has been studied by several workers. Although, its prime role in pathogenesis of
Cited by others 4 coronary heart disease has been consistently found, researchers have reported association of plasma/serum lipids
and lipoproteins with different cancers.[1],[2],[3],[4] The alterations in the circulatory cholesterol levels have
Total downloads in last been found to be associated with etiology of breast cancer and colorectal cancer.[5],[6],[7] However, only a few
2 months reports are available on plasma lipid profile in head and neck cancer.[1],[8] Head and neck cancer is one of the
Sep leading causes of morbidity and mortality due to cancer. Its incidence is much higher in Asian countries, as
16
2010
compared to the West. At our Institute (The Gujarat Cancer and Research Institute), head and neck cancer
Aug
76 accounts for about 2300 new cases every year which is around 25.3% of all cancer cases.[9]
2010

Ads by Google
Mouth Cancer The habit of tobacco consumption is a known etiologic factor for development of oral precancerous diseases and
Tongue Cancer head and neck cancer.[10],[11] Patients with oral precancerous conditions (OPC) have also been reported to
Throat Cancer
Lung Cancer
show a significant tendency to develop cancer.[12] It is believed that tobacco carcinogens induce generation of
Breast Cancer free radicals and reactive oxygen species, which are responsible for high rate of oxidation/peroxidation of
polyunsaturated fatty acids. This peroxidation further releases peroxide radicals. This affects essential
constituents of the cell membrane and might be involved in carcinogenesis/tumorigenesis.[13] Because of the
lipid peroxidation, there is a greater utilization of lipids including total cholesterol, lipoproteins and triglycerides
for new membrane biogenesis. Cells fulfill these requirements either from circulation, by synthesis through the
metabolism or from degradation of major lipoprotein fractions like VLDL, LDL or HDL. Earlier reports have
shown that antioxidant vitamins have protective effects against lipid peroxidation.[14],[15],[16] Lower blood
lipids have been associated with various cancers.[2],[4],[17],[18] Furthermore, some investigators have also
found relation of low serum cholesterol with increased risk of cancer occurrence[19],[20] and mortality.[21],[22]
The plasma concentrations of lipids are not the single additive function of intake, utilization and biosynthesis
because of its continuous cycling in and out of the blood stream. The question that whether hypolipidemia at the
time of diagnosis, is a causative factor or is a result of cancer has remained unanswered.

Considering these curiosities, the present study was aimed to evaluate the plasma lipid profile including: (i) total
cholesterol, (ii) LDL cholesterol (LDLC), (iii) HDL cholesterol (HDLC), (iv) VLDL cholesterol (VLDLC) and (v)
triglycerides in head and neck cancer patients, patients with OPC and healthy individuals. The association
between alterations in plasma lipid profile and habit of tobacco consumption was also studied.
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Translate » Material and Methods

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Subjects

Fifty-two healthy individuals, who had no complaint or any major illness in recent past, were included in the
study as controls. They were close relatives of the patients accompanying them during their hospitalization. One
hundred fifty three patients with OPC were also included in the study from the Government Dental College and
Hospital, Civil Hospital Campus, Ahmedabad. Out of these, 97 (63.4%) had oral submucous fibrosis, 44 (28.8%)
had oral leukoplakia and 12 (7.8%) were with other diseases including dysplasia and verrucous hyperplasia. The
hospital-based case control study included 184 untreated head and neck cancer patients, enrolled from the
outpatients' department of the Gujarat Cancer and Research Institute, Ahmedabad. The patients were diagnosed
as having cancer of oral cavity (n=121), pharynx (n=49), larynx (n=10) and maxilla (n=4). The diagnosis was
based on clinical and histopathological examinations. Staging was done according to UICC classification.[23]
[Table - 1] provides clinicopathological details of cancer patients. Histologically, 174 (94.6%) patients
represented with squamous cell carcinoma. Twenty-eight patients had early disease (stage I and II) and 131 had
advanced disease (stage III and IV). The classification according to their histological grade revealed that 64
patients had well differentiated, 85 had moderately differentiated, 20 had poorly differentiated and 5 had
undifferentiated carcinoma. The subjects in each group were further classified into as having no habit of tobacco
consumption (NHT) and with habit of tobacco consumption (WHT).

Blood sample collection

Fasting blood samples were collected from the subjects, into EDTA containing vacuettes. Samples were
centrifuged; plasma was collected and stored at -800C until analyzed.

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Methods
Plasma cholesterol levels were estimated using cholesterol kits obtained from Autospan Reagents, Span
Diagnostics (Gujarat, India). Briefly, 5 µl of plasma sample was mixed with 500 µl of working reagent that
contained cholesterol oxidase, cholesterol esterase, peroxidase, 4-amino phenanzone, surfactant, phenol, buffer,
preservatives and stabilizer. The mixture was incubated at 370C for 10 minutes and absorbance was read at 505
nm.
Plasma HDL cholesterol levels were also estimated using Autospan kits obtained from Span diagnostics (Gujarat,
India). Briefly, 0.3 ml plasma sample was mixed with 0.3 ml precipitating reagent (PEG 6000, stabilizer and
preservative), followed by 10 minutes incubation at room temperature. The mixture was centrifuged at 2000 rpm
for 15 minutes. The supernatant obtained was mixed with working cholesterol reagent. After incubation of 10
minutes at 370C, absorbance was read at 505 nm.
The triglyceride levels were estimated using liquid gold kits, obtained from Autospan reagents, Span diagnostics
(Gujarat, India). Briefly, 10 µl plasma sample was mixed with 1000 µl of triglycerides assay reagent containing
Pipes buffer, lipase, 4-chlorophenol, Magnesium ion, ATP, lipase, peroxidase, glycerol kinase, sodium azide,
4-amino antipyrene, glycerol-3-phosphate oxidase and detergents. The mixture was then incubated for 10 minutes
at 370C and absorbance was read at 505 nm. VLDLC and LDLC levels were calculated as shown below:
VLDLC = Triglycerides/5
LDLC = Total Cholesterol - (VLDLC) - (HDLC)
Statistical Analysis
Student's t-test was performed to compare mean values of the parameters. “P” value <0.05 was considered to be
statistically significant.
» Results
Habit of tobacco consumption in the subjects
Habit of tobacco consumption in one or the other form (smoking/chewing/snuff) is highly prevalent in this region.
As depicted in [Figure - 1], 95.4% of the patients with OPC and 85.8% of the cancer patients were consuming
tobacco in one or the other form. The details regarding habit of tobacco consumption are provided in [Table - 2].
Tobacco chewing in the form of readily available pouch (gutkha) and/or with lime was common in the patients
with OPC and bidi (country cigarette) smoking among the cancer patients. Habit of tobacco snuff (oral/nasal) was
usually observed in the female subjects, especially in female cancer patients. Use of tobacco, on an average, was
greater than 10 years in all the subjects. 79/107 (73.8%) of the smokers in cancer patients and 34/66 (51.5%) of
the smokers in patients with OPC had habit of smoking for more than 20 years. While, 29/72 (40.3%) of the
tobacco chewers in cancer patients had habit of chewing for more than 20 years.
Plasma lipid profile in the patients at the time of diagnosis
The mean plasma lipid profile of controls and patients, at the time of diagnosis, are represented in [Figure - 2]. A
significant decrease in plasma total cholesterol and HDLC were observed in cancer patients (P=0.008 and
P=0.000, respectively) as well as in patients with OPC (P=0.014 and P=0.000, respectively) as compared to the
controls. However, the mean plasma levels of total cholesterol and HDLC were comparable between patients
with OPC and cancer patients. The VLDLC and triglycerides levels were lower in cancer patients as compared to
the controls (P=0.059) as well as the patients with OPC (0.04). However, plasma levels of VLDLC and
triglycerides were comparable between controls and patients with OPC. Plasma LDLC levels did not reveal any
significant difference among all three groups.
Plasma lipid status in NHT and WHT subjects
Further, we compared plasma lipid levels between NHT and WHT subjects [Table - 3]. First, the plasma lipid
levels between tobacco consumers and non-consumers of the control group were compared. The controls
consuming tobacco showed lower mean plasma lipid levels than the non-consumers. Secondly, the plasma lipid
levels in tobacco habituates with OPC and cancer were also compared with the controls. Total cholesterol levels
were significantly lower in WHT subjects with cancer (P=0.02) and OPC (P=0.01) as compared to the NHT.
Likewise, HDLC levels were decreased in WHT subjects with cancer (P=0.001) and OPC (P=0.01) as compared
to the NHT. Significantly low level of HDLC in patients with OPC and cancer patients as compared to controls
(both NHT and WHT) [Table - 3]. When subjects were further categorized into above and below cutoff level
(considering median of NHT as cut off), it was found that greater proportion of the subjects with OPC and cancer
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were having total cholesterol, HDLC,VLDLC and triglyceride levels below cutoff limit [Figure - 3]

» Discussion

Cholesterol and triglycerides, important lipid constituents of cell, are essential to carry out several vital
physiological functions. Cholesterol is essential for maintenance of the structural and functional integrity of all
biological membranes. It is also involved in the activity of membrane bound enzymes and is important for
stabilization of DNA helix.[24] Cellular uptake and regulation of cholesterol is mediated by lipoprotein receptors
especially located on the surface of the cells. For transport in plasma, triglycerides and cholesterol are packaged
into lipoproteins, which are then taken up and degraded by cells to fulfill demands for cellular functions.

In some malignant diseases, blood cholesterol undergoes early and significant changes. Low levels of cholesterol
in the proliferating tissues and in blood compartments could be due to the process of carcinogenesis. The question
arises whether hypolipidemia is a predisposing factor or result of cancer? However, earlier studies have reported
that hypolipidemia may result due to the direct lipid lowering effect of tumor cells or some secondary malfunction
of the lipid metabolism or secondary to antioxidant vitamins.[21],[25]-[29] Several prospective and retrospective
studies have shown an inverse association between blood lipid profile and different cancers.[2],[4],[17],[18],[30]
Schatzkin et al[1] and Chyou et al[8] have observed an inverse trend between lower serum cholesterol and head
neck as well as esophageal cancers. The results are strengthened by the present study showing lower plasma total
cholesterol, HDLC, VLDLC and triglycerides in head and neck cancer patients and patients with OPC than the
healthy individuals.

Cholesterol is an essential constituent of lipoprotein fractions like LDL, HDL and VLDL. Seventy five percent of
the plasma cholesterol is transported in the form of LDLC. Body cells sequester cholesterol from LDL fraction of
lipoproteins. LDL receptors are necessary for metabolizing circulating LDLC levels and nearly 80% of the
plasma LDLC is cleared by LDL receptors.[31] High activity of LDL receptors attributes for lowering the serum
cholesterol levels. The individuals having deficient or defective LDL receptors remove plasma LDLC at much
lower rate and have considerably elevated levels.[32] In the present study, a significant decrease was observed in
plasma HDLC in cancer patients. This is in accordance with previous reports, that low HDLC is an additional
predictor of cancer[2],[30],[33] and it might be a consequence of disease that is mediated by utilization of
cholesterol for membrane biogenesis. We observed significantly decreased VLDLC and triglyceride levels in
cancer patients as compared to the controls and patients with OPC. Alexopoulos et al[18] have found
non-significant difference in serum triglycerides between controls and patients. While others have observed
elevated triglycerides levels in cancer patients.[2],[34]

Lipid peroxidation is an essential biochemical process that involves the oxidation of polyunsaturated fatty acids,
the important components of cell membranes. Tobacco carcinogens generate reactive oxygen species and lipid
peroxides, leading to tissue injury due to elevated lipid peroxidation, further damaging the cellular structural
blocks like lipids, proteins, DNA etc. Thus, lipid peroxidation may play a role in endogenous formation of
exocyclic DNA adducts. In our study, about 95% of the patients with OPC and 85% of the cancer patients were
tobacco consumers. Therefore, levels of the lipids in WHT subjects in patients with OPC and cancer patients
were compared with plasma lipid profile of NHT group of controls. Higher plasma cholesterol, HDLC, VLDLC
and triglycerides were observed in controls without habit of tobacco consumption than controls and patients with
tobacco habits. Earlier studies have shown an inverse relation between cholesterol and tobacco related cancers.
[1],[20] Neufeld et al[35] have reported passive smoking as a significant risk factor for decreased HDLC. Animal
studies have shown that nicotine, a known tobacco carcinogen, affects activity of enzymes responsible for lipid
metabolism.[36] It is also reported that LDLC uptake and cholesterol biosynthesis were decreased in the liver of
tumor bearing animals.[23] Further, exposure to tobacco carcinogens hampers antioxidant defense, leading to
accelerated lipid peroxidation. There is a strong relationship between vitamin E (a liposoluble antioxidant
vitamin) and lipids, especially cholesterol. Vitamin E is co-transported with all forms of cholesterol and
contributes to the first line of defense against lipid peroxidation.[14] Further, triglycerides and cholesterol are
positively correlated with vitamins.[37] We observed a significant decrease in plasma levels of fat-soluble
antioxidant vitamins such as vitamin E and b-carotene in tobacco habitués.[38] Together, our study shows
declined plasma levels of total cholesterol, HDLC, LDLC, VLDLC and triglycerides.

As majority of the patients with OPC and cancer patients were with habit of tobacco, marker levels in NHT
controls were compared with WHT controls, patients with OPC and cancer patients with habit of tobacco to
eliminate any bias due to tobacco habit. To establish a cutoff of circulating marker level equal numbers of
controls with and without habit of tobacco were enrolled. From the present study we can't predict the incidence
of tobacco habitués in healthy population.

In conclusion, Rose et al[39] reported 66% higher mortality rate due to cancer in the group of cancer patients
with lowest plasma cholesterol than in the highest plasma cholesterol. Our results add to this evidence of an
inverse relationship between lower plasma lipid profile and head and neck malignancies and oral precancerous
conditions. The lower plasma lipid status may be a useful indicator for initial changes occurring in neoplastic

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cells. However, a detailed study of cholesterol carrying lipoprotein transport and the efficiency of the receptor
system may help in understanding the underlying mechanisms of regulation of plasma cholesterol concentrations
in cancer. Further work on these interesting results as well as in-depth studies on lipoprotein transport and
receptor system is in progress in our laboratory.

» Acknowledgements

The authors are sincerely thankful to Cancer Treatment Research Foundation, Washington, D.C., USA, for
providing financial support (Grant No. G-96-134) for this study.

» References

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Figures

[Figure - 1], [Figure - 2], [Figure - 3]

Tables

[Table - 1], [Table - 2], [Table - 3]

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